
Class ^ (- 77 a 
Book TFs 



Copyright^ mS- 

CDPXRIGHT DEPOSHi 



THE 



Examination of the Urine 



OF THE 



HORSE AND MAN 



BY 



PIERRE A. FISH, D.Sc, D.V.M. 

PROFESSOR OF VETERINARY PHYSIOLOGY 

NEW YORK STATE VETERINARY COLLEGE 

CORNELL UNIVERSITY 



THIRD EDITION 
REVISED 



Comstock Publishing Co., Agents 

Ithaca, N. Y. 

1919 



SF7-5 
11 H 



Copyright 1919 

by 

PIERRE A. FISH 



Ukb 10 1919 



PRESS OF W. F. HUMPHREY, GENEVA*, ".. 



©CU536 984 



<-v % 



PREFACE. 



This manual represents the published form of mimeographed sheets, 
which the writer has used, for some years past, in giving instruction to his 
veterinary students. Data relating to the urine of the horse is not abundant 
and, unfortunately, some of the tests which may be standard for human urine 
are not reliable for that of the horse and, therefore, require modification. 
In order to emphasize this fact, the writer has employed the comparative 
method and asks the students to examine their own urine along with that of 
the horse and note carefully the differences in the results. 

The importance of urine examination for diagnosis and prognosis in 
human medicine is too well known to require emphasis here; but, notwith- 
standing certain difficulties in the way of its application in veterinary medicine, 
the writer believes that there is no important reason why just as much valuable 
information may not be derived, in certain cases, from the urine of the horse 
by an up-to-date veterinarian, as the physician obtains from the urine of his 
patients. 

Simplification of methods, without too great a sacrifice in accuracy, 
is essential for satisfactory examination, — especially by veterinarians. The 
writer realizes that only a short step has been taken in this direction, but 
hopes that time will lighten the difficulties. 

P. A. F. 

October, 1906. 



PREFACE TO THE SECOND EDITION 

In the present edition some changes have been made in the text and 
new material added with the hope of bringing the manual more completely 
up to date. There is still much to be done in investigating and simplifying 
the tests applicable to the urine of the domesticated animals and it is to be 
hoped that more research may be developed along this line. 

P. A. F. 

November, 1911. 



PREFACE FOR THE THIRD EDITION 

Experience has shown the desirability of making certain changes and 

some additions to the previous edition in order to bring this work up to date. 

Acknowledgement is gladly made to Dr. C. E. Hayden and former students 

whose suggestions and interest have been of much value 

P. A. F 
March, 1919. 



(3) 



TABLE OF CONTENTS. 



CHAPTER I. 

The Secretion of Urine pp. 7-12 

CHAPTER II. 

Quantity, Color, Transparency, Consistency, Reaction, 

Degree of Acidity, Specific Gravity pp. 12-19 

CHAPTER III. 

Qualitative tests: Inorganic Constituents, Water, 

Chlorides, Sulphates, Phosphates, Carbonates. . . .pp. 19-24 

CHAPTER IV. 

Organic Constituents: Urea, Uric Acid, Hippuric 
Acid, Creatinin, Mucus, Indican, Oxalic Acid, 
Acetone, Diacetic Acid, Urobilin, Leucin, Tyrosin, 
Phenol pp. 24-33 

CHAPTER V. 

Abnormal Substances in the Urine: Albumin, Sugar. . .pp. 33-41 

CHAPTER vi. " 
Bile, Blood, Melanine pp. 41-45 

CHAPTER VII. 

Quantitative Analysis, Centrifugal Method: Phos- 
phates, Chlorides, Sulphates, Uric Acid, Uricom- 
eter, Urea, Albumin, Saccharometer, Sugar, Ehr- 
lich's Diazo-reaction pp. 45-56 

CHAPTER VIII. 

Volumetric Methods : Chlorides, Phosphoric Acid, Sulphu- 
ric Acid, Relation of Constituents in Normal Urine pp. 56-59 

CHAPTER IX. 

Chemical Examination of Urinary Deposits, Acid 
Urate of Soda, Uric Acid, Cystine, Pus, Blood, Al- 
kaline Urates, Triple Phosphates, Calcium Oxalate, 
Calcium Carbonate, Pus, Blood pp. 59-62 

CHAPTER X. 

Microscopical Examination of Urine, Unorganized Sedi- 
ments : Uric Acid, Acid Urate of Soda, Oxalate of 
Lime, Hippuric Acid, Calcium Sulphate, Calcium 
Phosphate, Triple Phosphate, Urate of Ammonium, 
Cy stin, Leucin, Tyrosin. Organized Sediments: 
Epithelial cells; Glycogen cells, Amyloid bodies, 
Hyaline, Granular, Epithelial, Fat, Hemorrhagic 
and Waxy casts; Cylindroids; Blood and Pus cor- 
puscles; Spermatozoa; Mucus; Bacteria pp. 62-73 

Form for Urine Examinations, Procedure in Examining 

a sample of urine pp. 74-77 

Appendix, Formulae for Reagents pp. 77-80 



URINE ANALYSIS. 



APPARATUS FOR THE LOCKER 



1 dozen test tubes, 6 inch 

1 dozen test tubes, 5 inch 

1 Minim pipette 

1 Beaker, 10 oz. 

1 Graduate, 30 cc. 

1 Graduate, 250 cc. 

1 Flask 

1 Funnel, 1 % inch 

1 Funnel, 3 inch 

1 Watch glass 

1 Evaporating dish, 8 oz. 

1 Urin'rmeter 

1 Glass rod 

1 Thermometer 

1 Crucible, 8 cc. 



1 Piece wire gauze 

1 Piece absorbent cotton 

1 Box matches 

1 Test tube brush 

1 Test tube rack 

1 Test tube holder, wire 

1 Tripod 

1 Piece muslin 

1 Pack filter papers, 3 inch 

1 Pack filter papers, 6 inch 

1 Sponge 

1 Clay triangle 

2 Tin cans 

1 Copper water bath 
1 Towel 



Special apparatus, not found in the locker, may be obtained, when needed, 
by handing an order for it to one of the assistants. 



(5) 




Plate 1. Schematic Section of the Kidney 



1. Cut portion of the Kidney. 2. Pyramid. 3. Papilla of the Pyramid. 
4. Glomerulus, enclosed in Bowman's Capsule. 5. Bowman's Capsule. 6. 
Convoluted portion of Renal Tubule. 7. Loop of Henle. 8. Collecting 
Tubule. 9. Capillary net work, — second set; the first set is the Glomerulus. 
10. Openings of Collecting Tubules on Papillae, from which the Urine drops 
into the Pelvis of the Kidney. 



THE SECRETION OF URINE. 



The blood supply to the kidneys is an important consider- 
ation in the secretion of urine. At the outset, one is impressed 
by the fact that a relatively large artery supplies a relatively 
small gland. The blood pressure of the renal artery is nearly, 
if not quite, as great as that of the abdominal aorta, which means 
that about as much pressure is required to force the blood through 
the kidneys as is required to send the blood through the pedal 
extremities. The renal vein is also a relatively large vessel as 
compared with the size of the kidney or with the efferent vessels 
of other glands. The pressure within the renal vein is very 
low, practically as low as that in the posterior vena cava with 
which it connects. 

The artery, after entering the kidney, breaks up into branches 
which pass between the pyramids. At the junction of the cortex 
and medulla these branches form arches in the substance of the 
kidneys and from these arches branches run outward into the 
cortex to supply the glomeruli; other branches pass inward to 
supply the pyramids. Each glomerulus has its afferent and 
efferent vessel and of these the efferent is smaller. 

On issuing from the capillaries of the glomerulus, the efferent 
vessel soon breaks up into a second set of capillaries which sup- 
plies the uriniferous tubules; so that in this arrangement the 
blood goes first to the glomeruli and later supplies the tubules. 
The blood from the second set of capillaries is finally gathered 
up by small vessels which unite to form ultimately the renal 
vein. The glomeruli and the uriniferous tubules are the portions 
of the kidney actively concerned in the production of its secretion — 
the urine. 

The following points are therefore worthy of special notice; 
in the malpighian body the arterial blood in the glomerular 
capillaries is separated from the inside of the capsule of Bow- 
man by a thin layer of flattened epithelial cells only; two sets 
of capillaries exist, one set forming the glomeruli and the other 
supplying the uriniferous tubules, the blood supplying the tubules 

(7) 



must have first passed through the glomeruli, and is therefore 
more concentrated; in certain portions of the tubules short 
cylindrical epithelial cells are found which are comparable to 
true secreting cells found in other glands ; and finally, the smaller 
size of the efferent as compared with the afferent vessel fulfills 
a condition which retards the flow of blood through the glomerulus. 

The conditions are favorable for a high and variable pressure 
in the glomerulus and for a lower and more constant pressure 
in the second set of capillaries around the uriniferous tubules. 
On account of the resistance offered by a double system of capil- 
laries the blood pressure in the kidneys is kept relatively high. 

The changes in blood pressure may be observed upon the 



Ct Rett Of ^_ URiN€ 




Fig. 1 



1. Artery. 2. Glomerulus. 3. Capsule of Bowman. 4. Convoluted 
portion of Tubule. 5. Capillary net work. 7. Loop of Henle. 8. Collect- 
ing Tubule. 9. Opening of Tubule on Papilla. 



9 

kidney itself by means of an apparatus known as the oncometer. 
With each rise in the blood pressure the kidney swells, with 
each fall in pressure it contracts, and a tracing can be obtained 
very similar to an ordinary blood pressure tracing. The glomeru- 
lus suspended in its capsule is also influenced by these changes. 
When the capillaries are engorged with blood the glomerulus 
fills the capsule, when collapsed there is an evident space between 
the membranes of the glomerulus and the capsule. 

In 1842 Bowman advanced a theory relating to the pro- 
duction of urine which, while recognizing to some extent certain 
physical factors, also brought out the view that the urine is a 
secretion. 

In 1844 Ludwig advanced the theory, sometimes referred 
to as the mechanical theory, in which only physical processes 
were involved, i. e., filtration, diffusion and osmosis. It is true 
that in the capillaries of the glomerulus there is a high resist- 
ance because of the smaller size of the efferent vessel, there is, 
therefore, a higher degree of pressure in the glomerulus than 
in the capsule in which it is suspended. This inequality of 
pressure is favorable to filtration. As the result of any filtra- 
tion from the glomerulus some of the water of the blood with 
a certain proportion of dissolved substances would pass into the 
beginning of the uriniferous tubule. The effect of such a filtra- 
tion would render the blood remaining in the glomerulus and 
second set of capillaries more concentrated, and in the second 
set of capillaries in connection with the uriniferous tubules the 
essential elements of an osmometer would be obtained,- — an 
animal membrane formed by the delicate wall of the capillary 
and the wall of the tubule, upon one side of which there is a dense 
fluid, the blood, and upon the other a weak saline solution, condi- 
tions which are favorable to the processes of osmosis and diffusion. 

In this theory, as a result of the interchange of these fluids, 
some of the water passes from the tubule to the blood, making 
it less concentrated, and the products of retrogressive meta- 
morphosis, — urea and other organic substances, — pass from 
the blood to the tubule, forming urine. An objection in connection 
with the diffusion of urea has been raised in that it is a well-known 
fact that the urine contains a much greater amount of urea 
than does the blood and that it would be contrary to the laws 



10 




1. Artery. 
Glomerulus. 



of osmosis and diffusion for a fluid weak in urea (the blood) to 
pass through a membrane a substance which has accumulated in 
greater amount in a fluid (the urine) on the other side of that 

membrane. It should also be 
remembered that the blood 
is constantly moving and pre- 
sumably the fluid in the tu- 
bules is doing the same. In 
other words, it would seem 
that if the conditions indicate 
an osmometer, it varies from 
experimental ones in that the 
fluids on either side of the 
membrane are moving in a 
definite direction. The pro- 
teid constituents do not nor- 
mally leave the blood on ac- 
count of their well-known in- 
disposition to osmosis. 

One important fact, however, 
remains unaccounted for in 
Ludwig's mechanical theory, and that is, if the uriniferous 
tubules are stripped of their epithelial cells, as they often are 
in disease, urea and some other nitrogenous products are no 
longer, or only imperfectly eliminated, and become stored up 
in the blood and produce the condition known as uremia, al- 
though the conditions of an osmometer remain. It must, there- 
fore, be admitted that there is some direct or elaborating action 
on the part of the epithelium as originally suggested by Bowman, 
although under normal conditions, transudation, diffusion and 
osmotic processes may occur coincidently. 

The theoretical conclusions of Bowman have been con- 
firmed and extended by the practical researches of Heidenhain, 
who injected indigo-carmine into the blood of animals and found 
that it was promptly removed by the kidneys. These organs 
were removed at suitable intervals after the injection and care- 
fully examined under the microscope. In no instance did he find 
any of the indigo in the capsules of Bowman, but it was found 
abundantly in the cells lining certain portions of the tubules, and 
in the lumen of the tubules near these cells. Similar experiments 



Fig. 2 

2. Afferent Vessel. 3, 
4. Capsule of Bow- 
man. 5. Efferent Vessel. 6. Capil- 
lary network. 7. Uriniferous Tub- 
ule. 8. Vein. 



11 

with the urate of sodium showed that it likewise was secreted at 
the same place and in the same manner. 

Further investigations have tended to diminish the impor- 
tance of the "mechanical" factor and to develop a selective or 
secretory function for the cells, even including those of the 
glomeruli, by virtue of which the cells will permit certain sub- 
stances to pass through and prevent others, among the latter, 
albumin. The view is advanced that the passage of the fluid 
through the glomerulus is not a mere transudation, but a matter 
of selection also. 

The selective action of the renal cells is both qualitative 
and quantitative. As an example of the former, it is shown 
by experiment that if some egg albumin be injected into the 
blood, it is promptly eliminated by the kidneys. Egg albumin 
is not very markedly different from the serum albumin of the 
blood; both are indiffusible, but the renal cells recognize the 
former as a foreign substance and immediately separate it from 
the blood. The sugar, in normal quantity in the blood, although 
a diffusible substance, is not selected. Urea, which is also dif- 
fusible, but existing in the blood in much smaller amount than 
sugar, is selected and appears in the urine. Why this should 
be so it is difficult to explain, although it is a well-known fact 
that the sugar serves as a food for the tissues and is needed by the 
system, while urea is a waste product and would be detrimental 
to the system if not eliminated. 

That the cells exert a quantitative selection is shown by 
the fact that when sugar is present in the blood in excessive 
amounts, 3 parts per 1000 or over, the excess is promptly elimi- 
nated. 

Diet influences the reaction of the urine. A vegetable 
diet favors an alkaline reaction; a diet of flesh favors an acid 
reaction. The urine of the herbivora is therefore alkaline, while 
in the carnivora and omnivora the urine is mainly acid, although 
influenced to some extent by the kind of food eaten. The forma- 
tion of an acid fluid from alkaline material, — blood and lymph, — 
is at first sight puzzling. The well-known fact that the gastric 
secretion of all mammals is acid is a case in point. Further- 
more, experimental evidence shows that if an alkaline solution 
of sodium bicarbonate be placed upon one side of the membrane 
in an osmometer and a solution of neutral sodium phosphate be 



12 

placed upon the other side and a weak electric current be sent 
through the solutions, the fluid in contact with the positive pole 
will, in a short time, become acid from the formation of acid 
sodium phosphate, while the fluid in contact with the negative 
pole is increased in alkalinity NaHC0 3 +Na 2 HP0 4 = Na 2 C0 3 +Na 
H 2 P0 4 . 

While it may be true to a considerable extent that the kid- 
ney merely removes certain constituents from the blood and 
transfers them to the urine, it has been shown that the activity 
of the renal cells is required in the production of hippuric acid, 
— a new product, as hippuric acid is not present in the blood. 

In general, the theory explaining the secretion of urine, 
according to observed facts, is 'one which, while recognizing 
the process as partly physical, also requires some process of 
activity or elaboration on the part of the kidney itself. 



II. 

The urine of all mammals may be regarded, for the most 
part, as a solution of constituents derived from the metabolism 
of the tissues of the body- Some of these constituents, espec- 
ially the inorganic, may appear in the urine in the same form 
as they are taken into the body in the food, e. g., sodium chloride; 
others, especially the organic, represent decomposition products 
derived from the food or tissues, e. g., urea, creatinin, etc. The 
composition of the urine may, therefore, to some extent be regarded 
as an index of tissue activity, and the examination of this secretion 
is of considerable importance in clinical diagnosis and prognosis. 

That there is a relationship between the diet and the renal 
excretion is shown by the examination of the urine of the three 
great classes of animals grouped according to the food they 
eat; herbivora, omnivora and carnivora. Perhaps the first and 
most striking characteristic of the urine of each group is its 
chemical reaction. In vegetable feeders it is normally alkaline; 
in flesh eaters it is markedly acid, and in the omnivora it may 
be acid or alkaline according to the preponderance of the fleshy 
or vegetable material in the food. 

The relative proportion of phosphoric and carbonic acids 



13 

in the blood depends very much upon the composition of the diet. 
The chief mineral constituents of animal food are phosphates; 
those of vegetable foods carbonates. The blood of herbivorous 
animals is therefore rich in the latter, that of the carnivora in the 
former, and the acids formed in the tissues will form bicarbonates 
in the blood of the herbivora, and acid phosphates in that of 
carnivora. Acid phosphates hold earthy phosphates in solution, 
and bicarbonates dissolve earthy carbonates. Hence the urine 
of carnivorous animals is rich in phosphates of lime and magnesia, 
that of the herbivora in calcium and magnesium carbonate. 

In the practical treatment of this subject it is convenient 
to regard the horse as the type for the herbivora; man for the 
omnivora; and the dog or cat for the carnivora. In all cases 
throughout this work it is to be understood that, unless especially 
directed, the same tests are to be performed upon the urine of 
man and horse and a parallel record of such tests, for comparison, 
is to be kept upon the blank pages. 

In man, fresh, normal urine is a clear, golden-colored, trans- 
parent liquid, having a peculiar aromatic characteristic odor, 
and a bitter saline taste. 

In the horse, the urine is of a yellowish color when passed, 
but turns to a deep brown color upon standing for a time, due 
to the oxidation of pyrocatechin. It is more or less turbid and 
of a viscous character. Its odor is somewhat ammoniacal and 
strongly aromatic and more penetrating than that of man. 

The urine is chiefly a solution of urea and certain organic 
and inorganic salts; epithelial cells and mucus may also be 
held in suspension. Like milk and other animal fluids, the urine 
is not of constant composition. It is influenced by various 
factors, such as food, the amount of water or- other fluids taken 
into the body ; the temperature of the skin ; the emotions ; blood 
pressure, general or local; exercise; the time of day; age; sex; 
and medicines. 

Quantity. The amount varies considerably. In man, the 
quantity for twenty-four hours ranges from 1000 cc. to 2000 cc. 
In the horse, the average amount is about 3000 cc. to 4000 cc, 
although it may go as high as 7000 cc. or 9000 cc. In the ox a 
still greater quantity is secreted, the usual limits being from 
4500 cc. to 19000 cc. In the sheep, from 250 cc to 700 cc In 



14 

the pig it varies from 1200 cc. to 6000 cc. In the dog it varies 
from 200 cc. to 900 cc, depending upon the size of the animal. 

Color. The color ranges from pale yellow to brown. The 
normal color of urine is due to pigments probably derived from 
the coloring matter of the bile. 

Transparency. The urine of the horse is normally more 
or less opaque, that of man should be transparent at the time 
of passing. Many pathological urines, however, are perfectly 
clear. Pathological turbidity may be due to urates, phosphates, 
pus, bacteria, spermatozoa, fatty globules, blood, etc. All 
urines become turbid after standing for a time. 

The urine of the horse is especially turbid at the end of 
micturition, exceptionally it may be clear but becomes turbid 
on cooling. Its opacity is due to the presence of earthy salts 
of the carbonate of lime precipitated and formed by the dis- 
engagement of a certain amount of carbon dioxide from the 
bicarbonate of lime. The turbidity increases when the urine 
remains for any length of time in the bladder; it reaches its 
maximum when the urine is cooled by exposure to the free air; 
it diminishes after the ingestion of a large quantity of water. 

A clear, limpid urine is generally pathological in the horse; 
it indicates polyuria and the reaction in this case is usually acid, 
exceptionally neutral or alkaline when the phosphates are modified 
qualitatively or quantitatively. 

The turbidity of horse urine is abnormal when it is due to 
the presence of the phosphate of lime, or of calcium sulphate, 
or acid salts; also from the existence of albuminoid substances, 
(exudates, leucocytes in interstitial nephritis). 

In other animals the passing of turbid urine is usually re- 
garded as abnormal. 

Consistency. In the horse the urine is very viscid on account 
of the contained mucus and sometimes of epithelial debris. It 
niters very slowly. Urine taken directly from the ureter or 
the pelvis of the kidney is still more viscid, having a consistency 
very similar to that of egg albumin and a specific gravity some- 
what higher than normal. 

Reaction. The reaction of human urine is acid, that of the 
dog more so than that of man. In the pig it is sometimes acid 
and sometimes^ alkaline, depending upon the diet. In the horse 



15 

and sheep it is alkaline, also in the ox, but in the calf and foal while 
suckling it is acid. Herbivorous urine is alkaline, but if such ani- 
mals are starved for a time they practically become carnivorous in 
that they are living upon their own tissues and under such con- 
ditions their urine becomes acid. 

In the herbivora the reaction is normally alkaline and is 
due to the presence of bicarbonates or carbonates of lime or 
potassium. It would appear that the salts present in the vegetable 
food undergo oxidation to form organic acids, and these in turn 
are transformed into bicarbonates or carbonates, causing alkalinity 
of the urine. 

In man the acidity is due to the presence of acid sodium 
phosphate NaH 2 P0 4 . The urine passed before breakfast and 
during fasting or perspiration is more acid than at other times; 
during digestion and after meals the acidity is decreased. 

The degree of acidity of the urine may be determined by 
the use of the acidimeter, a graduated glass tube devised by Dr. 
H. R. Harrower. His description of it and its use follows : 

"The acidimeter consists of a glass tube so graduated that 
10 cc. is the first measuring point. From this upward the tube 
is graduated in fifths of degrees up to 100°, each degree repre- 
senting the amount of decinormal sodium hydroxide solution 
required to neutralize 100 cc. of urine. The method of using 
the acidimeter is as follows : The tube is filled with the urine to 
be tested, until the lower edge of the meniscus is just on the 
10 cc. mark. Two drops of phenolphthalein indicator solution 
are added, and then with an ordinary medicine dropper deci- 
normal sodium hydroxide solution is slowly added, inverting 
the tube after each addition, until the color of the fluid has just 
.been changed from a yellow to a light rose pink. The acidity in 
degrees is now read off on the tube at the level of the fluid. The 
normal acidity of a mixed 24 hour specimen should be between 
30 and 40 degrees. 

(With very concentrated urines in which the acidity is 
above 100° the tube may be filled to the 5 cc. mark and water to 
the usual level. The resulting figures are, of course, doubled.) 

If the urine is alkaline in reaction and it is desired to esti- 
mate the degree of alkalinity decinormal hydrochloric or oxalic 
acid solution must be used in place of the sodium hydroxide, 
the pink color present being just discharged by the acid." 



16 

The "Acid Index" or "Acid Unit" may be obtained by 
multiplying the degree of acidity by the amount of urine passed 
in 24 hours. The normal man is about 40,000 acid units. 

In man there is increased acidity physiologically during the 
night; with a flesh diet; after strong muscular exertion; during 
the intervals of gastric digestion; after the ingestion of mineral 
acids. 

There is increased acidity, pathologically, in fevers; in rheu- 
matism; after asthmatic attacks; in emphysema, pneumonia 
and pleuritis. 

The urine is less acid, or alkaline, physiologically, during 
gastric digestion; after hot or prolonged cold baths; after pro- 
fuse sweating; after copious ingestion of vegetable acids and 
their salts. 

Pathologically in acute and chronic inflammation of the 
urinary tract as in cystitis; in decomposition of the urine in the 
bladder in retention; in some cerebral and nervous diseases: in 
anemia; chlorosis; and general debility. 

When the urine of man is set aside in a cool place it grad- 
ually becomes more acid. This is called acid fermentation. 
After longer exposure to a warm atmosphere the urine becomes 
neutral, and finally strongly alkaline in reaction. It becomes 
turbid, has an ammoniacal odor, and deposits triple phosphate, 
ammonium urate, and great numbers of microbes exist. This is 
alkaline fermentation and is due to the transformation of the 
urea into ammonium and carbon dioxide, by means of a ferment 
produced by an organism known as the micrococcus ureae. (Fig. 
20). This organism is said to be conveyed through the air and 
to exist commonly around the orifice of the urethra. As long 
as the urine is acid the organism does not exist in the bladder, 
but may sometimes gain entrance through the medium of a 
sound or catheter. 

Test the reaction of the urines with red and blue litmus 
paper. Some urines change both the red and blue paper 
and are termed amphoteric. (In taking notes of the experi- 
ments it is well to record the results in parallel columns, 
the horse urine in one column and the human urine in the 
other) . 

Specific Gravity. The specific gravity of human urine 
ranges from 1015 to 1025, the average being 1018 to 1020. That 



i: 



of the horse ranges from 1020 to 1050, the average being about 
1035. That of the cow is lower, ranging from 1015 to 1045. It 
seems to depend considerably upon the milk secreted. In milch 
cows the urine contains a greater amount of water and a lesser 
amount of solids. The specific gravity of the urine of the sheep 
ranges from 1015 to 1060; that of the pig from 1005 to 1025, and 
of the dog from 1016 to 1060, depending upon the diet. Cat, 
1020 to 1040. 

The specific gravity may be ob- 
tained in different ways. The sim- 
plest and most usual way is to employ 
the instrument known as the urin- 
ometer. Some urinometers are not 
strictly accurate, but they may be 
tested by filling the urinometer jar 
with distilled water at 15° C. (60° F.) 
Read the division of the scale corre- 
sponding with the surface of the 
fluid looking above or below the 
meniscus as is found to be the most 
correct for the zero reading. Always 
adhere to this method when using 
the same urinometer. Test the spe- 
cific gravity of the urines and make 
the necessary corrections. If the 
urine is warmer than 15° C. add 1 to 
the last right hand figure of the spe- 
cific gravity for every 4 degrees of C. 
temperature, or for every 7 degrees 
of extra F. temperature. As oppor- 
tunity presents, test the specific grav- 
ity of some warm, freshly passed 
urine. Test the same urine later, 
when cool, and note if any difference 
in the reading. 

Urinometers already corrected for the ordinary room tem- 
perature (70° F.) may be obtained, in which case the temper- 
ature corrections may be omitted. If the urine should be too 
dense to read easily on the urinometer, dilute it with an equal 
volume of distilled water and multiply the reading by two to 




18 

get the correct specific gravity. The variation of the specific 
gravity depends upon the amount of the solids in the urine. 
The amount of solids may be estimated with approximate accuracy 
from the specific gravity by Christison's formula (Hsser-Trapp's 
coefficient): "Multiply the last two figures of a specific gravity 
expressed in four figures by 2.33. This gives the quantity of 
solid matter in every 1000 parts." (The number of grams in 
1000 cc). 

Example. Suppose a patient passes 1400 cc. of urine in 
24 hours and the specific gravity is 1020, 20X2.33 = 46.6 grams 
of solids in 1000 cc. To ascertain the amount in 1400 cc. use 
the following proportion: 1000 cc. : 1400 cc. : : 46.6 grams is 
to X (65.24 grams). The total quantity of the solids of the 
human urine is about 60 grams for the 24 hours or approximately 
4%. 

The following method taken from the Alkalcidal Clinic may 
also be used: Multiply the quantity, in ounces, of the 24-hour 
urine by the last two figures of the specific gravity and this by 
1.1, the product will represent the total solids in grains. Thus, 
if the amount of urine voided in 24 hours be 36 ounces and its 
specific gravity 1021, the formula would be 36X21X1.1, equal 
to 831 grains, the normal amount for a person weighing 100 lbs. 
The amount for other weights may be determined by proportion. 
In general the amount of total solids is a measure (a) of 
the activity of tissue change ; (b) of renal integrity ; (c) of abnor- 
mal constituents in the urine. Hygienic conditions which favor 
increased metabolism, as abundant food, active exercise, etc., 
increase the solid matter in urine, while the opposite conditions 
decrease them. 

.With the urine normal or subnormal in amounts, the solids 
are deficient pathologically from defective and enfeebled meta- 
bolism as in senility; anemia as a result of syphilis, cancer, etc.; 
chronic alcoholism; functional or organic diseases of the liver; 
from renal failure as in acute nephritis; certain" conditions of 
chronic renal disease; at the close of Bright's disease; venous 
congestion of the kidneys, etc. With the urine increased in 
amount there may be a deficiency of solids in diabetes insipidus; 
interstitial nephritis; amyloid disease of the kidney; chronic 
parenchymatous nephritis. When ths urine is not increased in 
amount, the urinary solids are increased in fevers; lithemia; some 



19 

forms of dyspepsia. When the quantity of urine is increased 
the solids are increased in diabetes mellitus; phosphaturia ; 
azoturia (excessive secretion of urea) . 

Calculate the solids of the urines by the formulae previously 
given. After obtaining the result of 1000 cc. estimate the quantity 
in 1250 cc. of human, and 5450 cc. of horse urine. In record- 
ing the tests use parallel columns, one column for the horse 
and the other for the human urine. 



III. 

Qualitative Tests. In all cases the urines must be filtered 
and perfectly clear before attempting the examination. 

INORGANIC CONSTITUENTS. 

These consist chiefly of sodium, potassium, ammonium, 
calcium, and magnesium, combined with hydrochloric, phos- 
phoric and sulphuric acids. 

Water. The water of the urine is derived from the food 
and drink, a small quantity being formed in the body. It var- 
ies according to the activity of the sweat glands of the skin. 

Chlorides. Next to the urea the chlorides form the chief 
portton of the urinary solids. The chlorides are increased phy- 
siologically after the ingestion of salt foods and much water; 
mental and physical activities ; and during pregnancy. Pathologi- 
cally, they may increase after the crises of fevers; after 
the absorption of exudates ; in diabetes (occasionally) . 

The chlorides are decreased pathologically, in all acute 
fevers; pneumonia (often entirely absent during the height 
of the disease); in cholera; and in most chronic diseases. An 
increase, or the re-establishment of the excretion of chlorides 
in disease is generally a favorable sign. In pneumonia it is a 
precursor of the crisis, and may often take place before other 
symptoms reveal the favorable change. 

Test a portion of each urine with a few drops of silver 

nitrate solution. A white, cheesy or curdy precipitate 



20 

insoluble in nitric acid indicates the presence of silver chloride. 
The phosphate of silver may also be thrown down but the 
nitric acid dissolves it, keeping it in solution. 

Evaporate carefully a few drops of urine upon a glass 
slide, with a gentle heat over the flame. Octahedral or 
rhombic crystals may form, — a compound of sodium chloride 
and urea. Examine with the microscope. (Fig. 5). 

Sulphates. The sulphates are chiefly those of sodium and 
potassium. Only a small amount of them enters the body with 
the food, so that they are chiefly formed from the metabolism 
of proteids in the body. The above are known as ordinary 
sulphates. Another class known as the ethereal sulphates also 
exist. The proportion exists in the ratio of 10 of the ordinary 
to 1 of the ethereal in man. In the horse the proportion is about 
2 of the ethereal to 1 of the ordinary. The ethereal sulphates 
are formed by the combination of sulphuric acid with organic 
bases such as phenol, skatol, etc., which originate from putre- 
factive processes in the intestine. The amount of ethereal sul- 
phates is of importance in determining whether or not the diges- 
tive processes are going on normally. In general the sulphates 
are increased physiologically by the ingestion of sulphur and its 
compounds ; nitrogenous food ; and conditions of increased meta- 
bolism. 

After acidulating the urine with hydrochloric acid 
to prevent the precipitation of phosphates, add to a small 
part of each urine, a little 2% barium chloride solution; 
a precipitate of barium sulphate is formed, insoluble in 
nitric acid. 

To separate the ethereal sulphates, mix 30 cc. of urine 
with an equal bulk of "baryta" mixture. Stir and filter. 
This removes the ordinary sulphates (as barium sulphate), 
add 10 cc. of hydrochloric acid to the above filtrate, and 
keep in the water bath at 100° C. for an hour in the hood 
and then allow the ethereal sulphates to settle. This may 
require some little time. (Baryta mixture is prepared by 
making saturated solutions in the cold, of barium nitrate 
and barium hydrate, and adding two volumes of the hydrate 
to one volume of the nitrate) . 



21 

Phosphates. The phosphates consist of alkaline and earthy 
salts in the proportion of 2 to 1. The latter are insoluble in 
an alkaline medium and are precipitated when acid urine becomes 
alkaline. They are insoluble in water, but soluble in acids; in 
acid urine they are held in solution by free C0 2 . The alkaline 
phosphates (sodium and potassium) are very soluble in water, 
and they never form urinary deposits. The earthy are phos- 
phates of calcium (Ca 3 P0 4 ) 2 (abundant) and magnesium (MgHP0 4 
plus 7H 2 0) (scanty). An alkaline medium precipitates them al- 
though not in the form in which they occur in the urine. 

The excretion of phosphates in the urine is largely dependent 
upon the amount of calcium ingested; the more calcium the 
food contains, the less phosphoric acid appears in the urine, and 
the more in the feces. This is due on the one hand to the tendency 
on the part of calcium to form insoluble calcium phosphates in 
the intestinal tract and in this way to prevent the absorption of 
the food phosphates; on the other hand, to the well-established 
tendency of calcium salts to be excreted into the bowel and not 
into the bladder; one must imagine in the latter case that calcium 
salts circulating in the blood combine with circulating phosphoric 
acid and bear the latter with them into the bowel. 

The fact is . of some therapeutic importance in the treat- 
ment of nephrolithiasis due to uric acid calculi, for the admin- 
istration of calcium salts in this affection by bearing much phos- 
phoric acid into the bowel, leads to the excretion of less phosphoric 
acid in the urine, and hence of normal and basic instead of acid 
phosphates; and as the latter precipitate and the former dissolve 
uric acid, it will be seen that by giving calcium we prevent the 
precipitation of crystalline uric acid and urate deposits in the 
urinary passages. (Croftan). 

Where considerable calcium is present in the food the excre- 
tion of phosphates in the urine is minimum, especially if the urine 
is alkaline because of the presence of sodium or potassium salts. 
In herbivorous animals where the urine is alkaline and where 
considerable quantities of phosphorus containing food are eaten, 
very little phosphate is excreted in the urine. 

Phosphates are increased in the urine pathologically in 
rickets; osteomalacin ; osteoporosis; fractures; chronic rheu- 
matism; diseases of the nervous system; and after great mental 



22 

strain and worry. Phosphates are decreased in renal diseases 
and phthisis. 

Physiologically, variations occur chiefly from the character 
of the food and drink; in the horse there may be an increase 
in the urinary phosphates after a large feed of oats, bran, oil- 
cake, etc. Pathologically, they are increased during the active 
changes in such bone diseases as spavin, ring-bone, splint, 
etc. 

To a small amount of each urine add about half its 
volume of nitric acid and then add two volumes of a 5% 
solution of ammonium molybdate and boil. A canary 
yellow ppt. (crystalline) of ammonium phospho-molyb- 
date should appear in the omnivorous urine. In the herbivor- 
ous urine the presence of so much organic matter renders the 
test unreliable; although a precipitate may form it is not a 
typical or characteristic one for phosphates in the urine of 
the horse, unless the organic matter has been previously 
removed. 

To each of the urines add half its volume of ammonia 
and allow it to stand. A precipitate of earthy phosphates 
is formed, in the urine of man Filter, add enough nitric 
acid to give an acid reaction to # the urines, and test the filtrates 
with ammonium molybdate as before. This method separates 
the earthy from the alkaline phosphates. 

To a portion of the urines add half their volumes of 
baryta mixture; a copious precipitate. Filter, add nitric 
acid and test the filtrates with ammonium molybdate. No 
ppt. should occur as the baryta mixture precipitates the 
phosphates as well as the sulphates and carbonates. 

Use a little of the magnesia mixture instead of the baryta 
mixture. Filter, add a little nitric acid, and test the filtrate 
with ammonium molybdate. (The magnesia mixture is 
composed of magnesium sulphate 1 part, ammonium chloride 
1 part, ammonia water 1 part, and distilled water 8 parts). 
To portions of the urines add a few drops of acetic 
acid and then a little 5% uranium nitrate solution, — a 
yellow ppt. of uranium phosphate is formed. 
The lime, magnesia, iron and other inorganic urinary consti- 
tuents are comparatively unimportant, and have no special 



23 



clinical significance. The tests for them are somewhat compli- 
cated and are therefore omitted. 

Demonstration of Carbonates and C0 2 in Urine. Carbon 
dioxide exists in the urine to some extent in a free state. There 
are also various carbonates present, especially in the herbivorous 
urine. The amount is very variable and to a great extent is 
dependent on the kind of food that is eaten; large quantities of 
vegetable foods determine an increase both in the combined and 
in the free carbon dioxide. The carbonates may be broken up 
and C0 2 given off by the application of heat or certain acids. 

Heat experiment. Fill a small flask about half full 
of unflltered herbivorous urine. Through the perforated 
stopper of the flask pass some bent glass tubing connected 
with a test tube containing lime water. Heat the urine 
in the flask, and as it boils the C0 2 will pass over into the 
test tube, and calcium carbonate will be formed from the 
union of the gas with the lime. Repeat the experiment 
with omnivorous urine. 

, A simple proof of the presence of carbonates, or C0 2 
in the urine is to fill a Doremus Ureometer with 25 cc. of 
the unflltered urine and introduce 1 cc. of nitric acid. The 
larger part of the gas (C0 2 ) rises and collects in the upper 
portion of the ureometer. Compare the amounts thus 
obtained in the urines of horse and man. 

A simpler qualitative test is to add a few drops of nitric 
or acetic acid to a little unflltered urine in a test tube. If 
effervescence occurs it is due to C0 2 set free from the car- 
bonates by the acid. 



24 

IV. 

ORGANIC CONSTITUENTS. 

Urea is, in amount, the principal constituent of the solids 
of the urine. It is the most important product of the decomposi- 
tion of proteid in the food. In round numbers it forms from 
2% to 3.5% of the urine, averaging about 2.5%. About one-half 
of the total solids in the urine consists of urea. Urea has no effect 
on litmus, it is odorless, has a weakly cool and bitter taste like 
saltpeter. It is very soluble in water amd alcohol, but it is almost 
insoluble in ether and benzine. About 90% of the total nitrogen 
of the urine is excreted in the form of urea. 

Physiologically, urea is increased by a proteid diet; exercise 
and muscular vigor ; by drinking much water. It is decreased by 
fasting; non-nitrogenous food; reduction of water in the diet; 
alcoholic beverages, tea or coffee; indolence of mind and body. 
Pathologically, it is increased in all acute fevers; dyspnoea; 
diabetes; and phosphorus poisoning. It is decreased in uremia; 
acute yellow atrophy of the liver and in chronic diseases. In 
general, any disease that interferes with the activity of the liver 
decreases the urea. Any disease affecting the uriniferous tubules 
may modify the appropriation of the urea from the blood and 
affect its passage into the urine. An increase of the urea inde- 
pendently of the physiologic variations and amount of nitro- 
genous food eaten is an approximate index of the amount of 
tissue waste in the system; on the other hand, when the urea is 
decreased it is an evidence of a diseased condition of the liver 
(the producer of urea) or the kidney (the eliminator of urea). 
A simple method of detecting urea is to concentrate 
a small amount of dog* or human urine in an evaporating 
dish to about half of its original volume. Place a drop 
or two of this concentrated urine upon a glass slide and 
after adding a drop of nitric acid, gently warm over the 
flame. If urea be present, upon evaporation, the micro- 
scope will show the characteristic crystals of nitrate of 
urea, of rhombic or hexagonal form. (Fig. 4). 

*For this experiment, dog urine is more satisfactory than human because 
of the larger percentage of urea present. 



25 



Take 20 cc. of fresh, filtered dog or human urine and 
add 20 cc. of baryta mixture to precipitate the phosphates 
and sulphates. Filter, evaporate the filtrate to dryness, 
and extract the residue with a little boiling alcohol over 
the water bath very carefully. Filter off the alcoholic 
solution, stand the.filtrate away in a cold place for crystalliza- 
tion. The crystals of urea, usually long, fine, transparent 
needles, will separate out. Examine them under the micro- 
scope. (Fig. 5). 

Repeat in the hood, the experiment upon the urine of 
the horse and compare results. 

Heat some urea crystals in a test tube. Biuret is 
formeTI and ammonia comes off. Add a trace of copper 
sulphate solution and a few drops of 20% caustic potash. 
A rose-red color is produced, — the biuret reaction. 





Fig. 4 

Crystals of Nitrate of Urea. 



Fig. 5 
Crystals of Urea. 



Medicines which increase the amount of urea are: urea 
itself; uric acid, common salt, phosphoric acid, squill, theo- 
bromine, colchicum, cubebs, atropine, cantharides, vegetable 
acids, iron preparations, hyposulphite of soda, potassium chlor- 
ide, ammonium chloride, coca, potassium permanganate, oxygen, 
salicylic acid. 

Medicines which decrease the amount of urea are: digitalis, 
alcohol, coffee, tea, potassium and sodium iodides, potassium 
bromide, arsenic, turpentine, aklaline carbonates, mercury, anti- 
pyrin, valerian, quinine sulphate, benzoic acid. 



26 

Uric Acid, sometimes called lithic acid, is, next to urea, 
the most important nitrogenous constituent of the urine of man. 
It has been said to be absent in herbivorous urine, being replaced 
by hippuric. This has been shown by later researches not to be 
strictly true, as a trace of uric acid is found in addition to the 
hippuric. In birds and reptiles uric acid is the chief nitrogenous 
constituent, being present in greater amount than urea. 

In man the proportion of urea to uric acid is about 45 to 
1, about 0.5 gram of the latter being excreted in the 24 hours. It 
causes the brick red deposit sometime seen in urine after standing 
for a time. Its solubility is very low, only 1 part being soluble in 
14000 cc. of cold water, and 1 to 18000 in boiling. Physiologically, 
it is increased and diminished proportionately with urea. Patho- 
logically, it is increased in indigestion; acute dropsies, rheumatic 
and catarrhal inflammations; after attacks of gout; cancer of 
the liver; in leucemia; in all disturbances of the circulation and 
respiration. Pathologically uric acid is decreased in chronic 
diseases generally; diabetes and polyuria; before paroxysms of 
gout; anemias; chronic rheumatism ; chronic diseases of spinal 
cord. 

Uric acid is generally in solution in the form of urates of 
sodium, potassium, ammonium, lime and magnesium. These salts 
are very easily decomposed even by weak organic acids. 
Perform the following tests upon both urines (filtered). 

In a conical glass, add 5 parts of hydrochloric acid 
to 30 parts of urine. Label and put in a cool place for 
24 hours. Red or brownish colored crystals of uric acid 
are deposited upon the sides of the glass, or form a pellicle 
on the surface of the fluid like fine grains of cayenne pepper. 
The brownish-red color is due to pigment (uroerythrin) . 

Murexide Test. To about 1 cc. of human urine add 
a little nitric acid; evaporate in a porcelain dish very care- 
fully to avoid charring. Cool and add a drop of dilute 
ammonia, a purple red color of murexide or purp urate of 
ammonia is formed. It turns bluer upon the addition of 
caustic potash solution. 

Dissolve a few crystals of uric acid in 10% caustic 
soda or potash. Add a drop or two of Fehling's Solution — 
or dilute cupric sulphate and caustic potash and heat — - 



27 



there should occur a ppt. which at first may be white and 
after a time turning green or reddish. 

(Fehling's Solution is put up in two bottles; one labeled 
A, the other B. In making tests, take equal parts of A and 
B and add the substance to be tested. See formula in appen- 
dix.) 

Schiff's Test. Dissolve a little uric acid in a small 
quantity of 10% sodium carbonate solution. With a glass 
rod, place a drop of silver nitrate solution on filter paper 
and then a drop of the uric acid solution so that the two 
drops partially overlap. A dark brown or black spot of 
reducing silver appears. 

Hippuric Acid, (C9H9NO3), is found especially in the urine 
of the herbivora, as the horse, ox, etc. In the urine of carnivora, 
and especially in that of man, it exists in but very minute quantity, 
usually about 0.5 to 1 gram being excreted in 24 hours. It 
dissolves readily in hot alcohol but is sparingly soluble in water. 
It occurs in man after the ingestion of certain vegetables, such as 
asparagus, plums, pears, and apples with their skins; a purely 
vegetable diet and from the use of benzoic acid, cinnamic acid, 
essence of bitter almonds, quinine, and analogous bodies. Hippuric 
acid normally seems to be derived chiefly from the husks or cuticu- 
lar structures of the food. 

Pathologically hippuric acid is increased in diabetes, chorea; 
jaundice and other liver complaints; and in the acid urine of 




0> ^ 



Fig. 6 
Crystals of Hippuric Acid. 




• Fig. 7 

Various forms of Hippuric Acid 
with triple phosphates. 



28 

patients suffering from all kinds of fevers. In testing for hip- 
puric acid the fresh urine should be used; if stale, benzoic acid is 
likely to be obtained instead. 

Test. Saturate the fresh urine with lime water, which 
transforms the hippuric acid into a salt of lime, the fluid 
is boiled, then filtered, evaporated to a syrupy consistency, 
cooled and excess of hydrochloric acid added, when hippuric 
acid crystallizes out on standing for 24 hours. The horse 
urine is to be evaporated in the hood. 

Creatinin (C4H7N3O). This substance was discovered in 
the urine by Liebig. It is easily produced from creatin, a sub- 
stance normally existing in plain and striated muscular tissue. 
Creatinin occurs constantly in normal human urine, the amount 
varying according to Voit, from 0.5 to 4.9 grams per day accord- 
ing to the quantity of proteids eaten. It is said not to be dimin- 
ished by fasting. 

Pathologically it is increased in 
typhoid fever; intermittent fever; 
pneumonia; tetanus. It is de- 
creased during convalescence from 
the above diseases ; and likewise 
in anemia; chlorosis; muscular 
atrophy; tuberculosis; paralysis, 
etc. 

Test. Take 5 cc. of urine in 

a test tube and add a few drops of 
Fir 8 
n . ' . freshly prepared 1 % sodium nitro- 

Creatmm. . f tl , ' , . „ 

prusside. Render the solution alka- 
line with potassium hydroxide. A ruby red color develops which 
soon turns yellow. The above is Weyl's test. Acetone, if pre- 
sent, also gives the red reaction. 

Perform the above test on both urines. 
Caution. Creatinin' reduces copper oxide and may 
be taken for small quantities of sugar. 

Mucus. Mucus in the urine is not visible, but causes cloudi- 
ness sometimes by entangling epithelial cells, urates, oxalate 
of lime and other crystals in various amounts. 

Add to the urine a little acetic or citric acid and, in 

addition a few drops of liquor iodi comp. (Lugol's solu- 




29 



tion) which makes the threads or bands of mucin visible. 
Indican or Indoxyl. This substance is derived from indol, 
one of the putrefactive products formed in the intestine. Indoxyl 
occurs in very small quantities in normal human urine, about 
.004 to .020 gram for the 24 hours. Horse's urine is said to con- 
tain 23 times as much. The intestines of the herbivora are much 
longer than in the case of the carnivora. On this account, and in 
conjunction with the carbohydrate diet, a much greater fermen- 
tation occurs, which leads to a greater elimination of indican in 
the urine. 

In obstruction of the intestine, or in intestinal catarrh or 
where the food remains a long time in the intestine and ferments 
there, the proportion of indican increases in the urine and causes 
a true indicanuria. Indoxyl is of considerable clinical import- 
ance, an increase is indicative of imperfect performance of the 
digestive processes. In obstructive diseases of the small intestine 
the increase of indoxyl in the urine is enormous. 

Pathologically indoxyl is increased in cholera, typhoid fever, 
peritonitis, dysentery, Addison's disease, cancer of the liver and 
stomach and pernicious anemia. 

Obermayer's Reagent. This reagent has the advantage 
of keeping indefinitely. It is prepared by dissolving 2 grams 
of solid ferric chloride in 500 cc. of concentrated hydro- 
chloric acid. (Sp. gr. 1.19). 

To equal parts of urine and the above reagent add a 
little chloroform. Shake frequently but not too violently 
(otherwise an emulsion may be formed). The chloroform 
will become more or less blue by the indigo formed, in pro- 
portion to the indican originally present. According to 
the depth of the blue color it may be designated as little, 
much or copious. 

A test said to be somewhat more sensitive is as follows : 
Urine 10 cc; 20% basic Lead acetate 2 cc. Shake and 
filter. Add to the filtered urine 10% Thymol in alcohol, 
yi cc; Obermayer's Reagent 10 cc. and let stand for 15 
minutes; add chlorofoon, 4 cc. Shake gently several times. 
Let the chloroform settle. A violet color indicates the pre- 
sence of indican. 

Jaffe's Test. To a little urine add an equal volume 
of strong hydrochloric acid. Add to this mixture 2 or 3 



30 



drops of a solution of freshly prepared chlorinated soda. 
There soon forms a bluish cloud of indigo. Add a little 
chloroform and shake gently, this will take the indigo into 
solution and settle as a blue layer at the bottom of the test 
tube. The amount of indoxyl can be judged by the depth 
of the blue color. Indican is oxidized by free chlorine ob- 
tained from the chlorinated soda to indigo. 

Oxalic Acid. This is usually found in combination with 
lime in the form of calcium oxalate. The crystals are of small 
size and appear in the form of dumb bells and octahedra. They 
occur normally in greater amount in herbivorous than in omni- 
vorous urine. They greatly increase after eating such vegetables 
as tomatoes, fresh beans, beet-root, asparagus, apples, grapes, 
honey, and after the use of rhubarb, senna, squills, etc. Another 
source of oxalic acid in the body is incomplete oxidation of carbo- 
hydrates and proteids or retarded metabolism. It is therefore a 
result of mal-assimilation and is found in dyspepsia, diabetes mel- 
litus, etc. The long continued excretion of an excess of oxalate of 
lime frequently irritates the kidneys, producing albuminuria and 
grave nervous disturbances and may lead to the formation of 
calculi. (Fig. 18.) 

Acetone. Normal urine may contain traces of acetone but 
it occurs in excessive quantities as a pathological condition.* It 
is found in many of the fevers, certain forms of cancer, in starva- 
tion, and in diabetes, when it indicates an advanced form of the 
disease. It is associated with an increased proteid metabolism 
and is looked upon as a product of proteid decomposition with 
deficient oxidation. 

Lieben-Ralfe Test. Dissolve 1.3 grams (20 grains) 
of potassium iodide in 4 cc. (1 dram of liquor potassae). 
Boil in test tube, after which gently pour the urine on its 
surface. A yellow precipitate between the two solutions 
indicates an affirmative test. A more satisfactory test 
is to add to the urine a few crystals of iodine and of iodide 
of potassium with some caustic potash. Heat. Yellow 
precipitate — iodoform with its characteristic odor. 

Legal' s Test for Acetone. Add to 5 cc. of the urine 

*If pathological urine is not available, a small amount of acetone may 
be added to the urine for laboratory tests. 



31 



some fresh aqueous solution of sodium nitroprusside fol- 
lowed by a little ammonia, or sodium hydrate solution, 
which gives a red color. Add an excess of acetic acid and 
if acetone is present the red color will be intensified, if acetone 
is absent a yellow color will result. Compare with creatinin. 

These tests are not always satisfactory when applied 
to the ordinary urine. Greater accuracy is claimed if the 
urine is distilled and the tests applied to the distillate. 

Frommer's Test for Acetone. To 10 cc. of urine in 
a test tube add some potassium hydroxide to make the urine 
markedly alkaline; before the latter is dissolved add 10 to 12 
drops of salicylic aldehyde (made by dissolving 1 part of 
salicylous acid in 10 parts of absolute alcohol). Heat the 
mixture to about 70° C. With acetone the solution becomes 
yellow, then red and after long standing dark red. According 
to Frommer, even the minutest amount of acetone will give 
this reaction and no other constituent of the urine will give 
this color — not even diacetic acid. The reaction is explained 
as follows: One molecule of salicyclic aldehyde combines 
with one molecule of acetone to form oxybenzol-acetone. 
This, in the presence of strong alkalies, forms dioxy-dibenzol- 
acetone. The alkaline salts of this compound are intensely 
red. 

Diacetic Acid. Lindemann's modification of Riegler's test. 
Urine 10 cc. 

Acetic Acid 5 drops 

Lugol's Solution 5 drops 

Chloroform 3 cc. 

With normal urine the chloroform is colored rosy red; with 
urine which contains diacetic acid it remains colorless. Urine 
which contains much uric acid should have the amount of Lugol's 
Solution doubled, and should not be too vigorously shaken. 

Urobilin is commonly regarded as the most important color- 
ing matter in the urine. There is some evidence that it represents 
a reduced form of bilirubin, one of the pigments of the bile.* 
Urobilin is more readily obtained from highly colored urines, 
(fevers, etc.)'. 



*A small amount of an alcoholic extract of the feces added to the urine 
will usually give favorable tests for urobilin. 



32 



Test. The ordinary test with an alcoholic solution 
of zinc may be simplified in the following manner: 10 cc. 
of urine are acidified with 2 drops of HC1 and shaken with 
2 cc. of chloroform. After the separation of the liquids, 
2 cc. of the chloroform layer are tested with 4 cc. of a solution 
of 1 gram of crystallized acetate of zinc in a liter of 95% 
alcohol (shelf reagent). At the junction of the two layers 
the green fluorescent ring characteristic of urobilin will 
appear,- and on shaking, a fluorescence which is rose-colored 
by reflected light will be distinguished throughout the liquid. 

Another test is to add ammonia to the urine until dis- 
tinctly alkaline, filter, and to the filtrate add a little 10% 
chloride of zinc solution. A green fluorescence should 
appear, and if examined with the spectroscope, a characteristic 
band should occur. (See Fig. 16.) 

Leucin and Tyrosin are pathologic constituents of urine. 
They are normal products of pancreatic digestion and under 
ordinary conditions are carried, after absorption, to the liver, 
where they disappear, presently undergoing decomposition. 
When present in the urine these bodies are usually considered 
pathognomonic of acute yellow atrophy of the liver, although 
they are likewise stated to be present in the urine in certain 
rare cases of acute phosphorus poisoning associated with hepatic 
atrophy, due to typhoid fever, etc. 

Phenol. According to Tereg and Munk the horse excretes 
in the urine about 10 grams of tribromphenol in 24 hours. The 
tribromphenol is equivalent to 3 grams of phenol daily. Great 
importance is laid by these observers on the excretion of phenol, 
a process which is suspended during intestinal complaints, par- 
ticularly colic, and is, according to them and others, a cause of 
the rapid death in these effections, produced by the toxic effect 
of the unexcreted phenol. The production of phenol in the 
healthy body is greatly influenced by diet, being largest on rye 
and hay, one part peas and two parts oats, and on hay alone; it 
is smallest on rye alone, and next smallest on oats and hay. Sal- 
kowski is inclined to regard the excretion of 3 grams of phenol 
daily as too high. 



33 



V. 
ABNORMAL SUBSTANCES FOUND IN THE URINE. 

Albumin. The presence of this substance in the urine is 
regarded as pathologic. There is, however, in the urine of some 
individuals apparently enjoying perfect health, minute traces of 
albumin sometimes present, and, unless these traces persist, are 
not to be regarded as serious. If present in any considerable 
quantity, it must be regarded as distinctly abnormal. Albumi- 
nuria is the term applied when albumin occurs in notable quantity 
in the urine. The principal form of albumin present is serum- 
albumin, in addition there may be serum-globulin, acid albumin, 
albumose, and peptone. 

The amount of albumin in the urine may be increased: 1. 
By food rich in albumin; 2. Suppression of cutaneous perspira- 
tion, as by colds, burns, or cutaneous diseases; 3. Pulmonary and 
cardiac diseases attended with dyspnoea, cyanosis, valvular dis- 
eases of the heart; 4. Febrile and inflammatory diseases, as ma- 
larial, eruptive, typhus and typhoid fevers, croup, diptheria, 
erysipelas, rheumatism, gout, peritonitis, meningitis, etc.; 5. By 
lesions or prostration of the nervous system, especially when at- 
tended by diminished temperature and arterial tension, as from 
grief, fear, injury, pressure; 6. Pressure as from tumors, preg- 
nancy, etc. ; 7. Cachexias, as from cancer, syphilis, scrofula, sep- 
ticemia; 8. Hydremia and ailments that disturb the vascular ten- 
sion; 9. Chorea, convulsions, exacerbations of febrile and other 
diseases; 10. Diseases of genito-urinary organs, as Bright's dis- 
ease, cystitis, hemorrhage, abscess, etc.; 11. Medicines, such as 
copaiba, cubebs, turpentine, some emetics and drastic cathartics, 
some anesthetics, coffee, many metallic salts, poisoning by hydro- 
gen arsenide, carbon protoxide, carbon dioxide, phosphorus, 
iodine, iodoform, etc. 

The loss of organic material (albumin) disturbs nutrition, 
the blood becomes more aqueous ; it sometimes produces an anas- 
arca which is the result of hydremia and of anuria. 

Albuminuria may be caused: 1. By an alteration in the renal 
transudation; 2. By certain changes in the blood; 3. By disturb- 
ance of the circulation. 



:34 



Renal Changes. Lesions of the dialysing portions of the 
kidney, especially of the glomerule; the epithelium of the convo- 
luted tubules may also be essential in the production of albu- 
minuria. These cells normally prevent the albumin from filtering 
through with the other elements of the plasma. Albuminuria is 
also dependent upon renal lesions, sometimes primary as in 
nephritis, sometimes consecutive as in alteration in the blood or 
a disturbance in the circulation. Acute nephritis, chronic neph- 
ritis, fatty or amyloid degeneration interfere with the process of 
dialysis. 

Bacteria may exercise a traumatic action upon the renal 
epithelium and cause desquamation or degeneration, obstruct the 
vessels, modify blood pressure, or by the excretion of soluble 
products irritate the parts. 

Changes in the Blood. The dialysing membrane cannot 
stand with impunity any adulteration of the blood. The passage, 
in the kidney, of any such substance as biliary pigment in icterus; 
glucose in diabetes; poisons, such as alcohol, lead or mercury, or 
toxic gases, render the urine albuminous. 

Subcutaneous injections of solutions of extractives (leucin, 
tyrosin, creatinin, xanthin and hypoxanthin) cause degeneration 
of the epithelium of the kidneys and albuminuria. The subcu- 
taneous injection of tincture of cantharides causes, in a few min- 
utes, the production of an albuminous exudate in the glomerules. 
Although the limit of saturation of the blood plasma by albumin 
may be unknown, it is none the less evident that a superabun- 
dance of the substance (albumin) in the vessels causes albuminuria. 

The existence of a physiologic albuminuria is still doubtful in 
the domestic animals; for Frohner, who has examined the urine 
of a number of healthy horses, has found only two cases in which 
albumin was present. In man it has been demonstrated to be due 
to severe muscular exercise, slight cold, and nitrogenous diet. 

Disturbance of Circulation. Too great a variation in blood 
pressure will cause albuminuria. 

Renal emboli, section of \-aso-motor nerves of the kidney, 
medullary lesions, etc., cause albuminuria by causing an active 
congestion of the kidney. Venous stasis, organic affection of the 
heart, of the liver, presence of fetus, etc., cause albuminous urine. 

The urine may be less fluid. Bacteria may frequently cause 



35 



thromboses, emboli, edemas, anemias, etc., from vaso-motor trou- 
bles changing simultaneously the filter, the liquids which filter, 
with regard to pressure and velocity. 

Albuminous urine is usually of light color and low specific 
gravity. It may occasionally be dark and dense, due to other 
ingredients, or to concentration. 

Under particular conditions of fatigue or disease albumin 
may appear in the urine. 

Temporary albuminuria is sometimes induced by a cold 
bath, especially in persons prone to kidney disease, and it has 
been observed after excessive muscular exercise, as in the urine 
of soldiers after a prolonged march. 

Any cause which leads to an increased blood pressure in the 
kidneys tends to induce albuminuria, and many of the cases in 
which it is the result of disease may be traced to this cause. Al- 
buminuria is a constant accompaniment of the nephritis follow- 
ing scarlet .fever and may occur to a less extent in pneumonia, 
typhoid and diptheria. It may also occur in diabetes, and is 
then a highly unfavorable symptom. 

In every case the urine must be clear before testing, by 
filtering it carefully; also take the specific gravity.* In addi- 
tion to the suspected urines make control tests on the normal for 
comparison. 

Heat Test. Heat about 5 cc. of the urine to the boil- 
ing point in a test tube. Note the slightest turbidity. If 
present it will be due to albumin or earthy phosphates. In 
horse urine the precipitate may be due to driving off C0 2 
and precipitation of lime, etc., not phosphates. Add slowly 
a few drops of acetic acid (or nitric). If due to the phos- 
phates the urine becomes clear, if the turbidity remains 
it is albumin. Care must be taken, in the addition of the 
acid after boiling, to note the effect after each drop is added 
and to go on adding until there is no doubt that the urine 
is distinctly acid. If only a trace of albumin is present 
and too much acid is added the albumin may be converted 
into acid albumin and remain in solution. Heat does not 
coagulate acid albumin. Add a little acetic acid to dissolve 
any phosphates and heat again. 

*If a pathological urine is not available, a little blood serum added to 
the normal urine will give satisfactory tests. 



36 



Another Heat Test. Fill a test tube about one-third 
full of water and boil it. Add a few drops of the suspected 
urine. If albumin is present a cloudiness or coagulum will 
appear, according to the amount of albumin present. 

Heller's Cold Nitric Acid Test. Pour some of the 
urine gently upon the surface of some nitric acid in a test 
tube. A ring of white coagulum occurs at the junction of 
the two fluids. If the quantity of albumin is small, the 
coagulum may not occur for a few minutes. A brown zone 
will frequently be seen at the point of contact due to the 
action of the acid upon the coloring matters of the urine, 
but it does not give any turbidity unless albumin be present. 

Millard-Robert's or Nitric Magnesian Test. The re- 
agent is as follows: Nitric Acid, 1 part; Sat. Sol. Mag- 
nesium Sulphate, 5 parts. Use as in the preceding test. 

Picric Acid Test. (Johnson's). Fill the test tube 
half full of urine. Slightly incline the tube and gently 
pour down its side about 2 cc. of a saturated solution of 
picric acid, so that it may come in contact with the upper 
layer of the urine. Place the tube in an upright position. 
A layer of coagulated albumin will appear at the line of 
junction. The coagulation of albumin takes place at once 
and is thus not easily mistaken for precipitated urates, 
which require some time for their precipitation and dis- 
appear on the application of heat. 

Ferrocyanide Test. To 2 cc. of acetic acid in a test 
tube add 4 cc. of a 5% solution of potassium ferrocyanide. 
Mix them and add 10 cc. of urine. A precipitate will appear 
if albumin be present. No heat is required. 

A test equally as good is that proposed by Zouchloss 
in which potassium sulphocyanide is substituted for the 
ferrocyanide. 

Sugar in the Urine. Dextrose or glucose exists in the blood 
from 0.8 to 1.25 parts per 1000. When a greater amount than 
3 parts per 1000 exists, the excess is excreted through the kidneys. 
It is maintained by many that a trace of dextrose is normally 
present in the urine and may appear in slightly larger quantities 
transitorily without pathologic significance. The presence of 
small quantities of sugar in the urine is designated as glycosuria; 



37 



in larger quantities it is known as diabetes mellitus. The former 
condition if habitual, is unnatural, and may terminate in the latter. 

These diseased conditions do not necessarily point to diseases 
of the kidneys or urinary organs, but rather to the liver. The 
kidney, in ridding itself of this product (dextrose), becomes 
irritated, and this irritation extends down the entire canal, and 
we thus have a real polyuria produced. 

Sugar, in sufficient quantity to react to ordinary tests, is 
found in the urine physiologically during pregnancy and lac- 
tation; in infants under two months old; in old persons living 
largely upon starchy and saccharine food. Pathologically in 
diabetes mellitus; in impeded respiration from pulmonary dis- 
eases; in impeded hepatic circulation (functional and organic 
diseases of the liver) ; in diseases of the central nervous system 
(general paresis, epilepsy, dementia, puncture of the fourth ven- 
tricle) ; in intermittent and typhoid fevers, by the action of cer- 
tain poisons, as carbon monoxide, arsenic, chloroform and curare; 
in abnormally stout persons. 

The persistent excretion of easily recognizable quantities of 
sugar constitutes diabetes. The quantity of urine is often enor- 
mously increased, as much as 10000 cc. being passed in 24 hours, 
by man. The specific gravity is high, varying from 1025 to 1050. 
The color is usually pale, from the dilution — not diminution — of 
the urinary pigments. 

The presence of albumin interferes with the tests for sugar 
and must, in all cases, be removed by the addition of acetic acid 
and heat. The urine, after being filtered, may then be used for 
the sugar tests. 

The particular property of glucose, which is utilized for 
its detection, is its action as a reducing agent — its disposition to 
absorb oxygen. In this property it differs strikingly from sac- 
charose or common cane sugar. 

Principle of the Copper Tests. If a little copper sulphate 
and an excess of a solution of caustic potash be added to a solution 
of glucose, a clear blue solution results. Without the glucose, 
the alkali would precipitate the pale blue cupric hydrate, Cu0 2 H 2 ; 
and if the mixture were boiled this blue precipitate would be 
reduced to a black precipitate of Cupric Oxide, CuO?. The 
clear blue solution containing glucose, however, when boiled, 
changes from transparent blue to opaque yellow, and speedily 



38 



deposits a yellow, ultimately red, precipitate of cuprous oxide CuO. 
When the quantity of sugar is large the change is immediate. 
When small the reaction takes a few minutes for its completion. 
Fehling's Solution. Solution A. 34.64 grams of pure crys- 
talline copper sulphate are powdered and dissolved in 500 cc. of 
distilled water. Solution B. Sodio-potassium tartrate (Rochelle 
Salts) 173 grams. Pure caustic potash 125 grams. Add enough 
distilled water to make 500 cc. When using take equal parts of 
Solutions A and B. (If diluted with 5-10 vols, of water the test 
is said to be more sensitive) . 

Place some Fehling's Solution in a test tube and boil 
it. If no yellow discoloration takes place it is in good condi- 
, tion. Add a few drops of the suspected urine and boil. 
If the mixture suddenly turns to an opaque yellow or red 
color, the presence of a reducing sugar is indicated. Normal 
horse urine probably because of its pyrocatechin usually 
changes the color of the copper solution, but this does not 
indicate sugar. 

Benedict's Modification of Fehling's Test. Greater 
delicacy and accuracy are claimed for this test. There are 
two solutions as in Fehling's. The first is prepared by 
dissolving 34.65 grams of cupric sulphate in a small aaiount 
of water and made up to 500 cc. The second by dissolving 
100 grams of anhydrous sodium carbonate and 173 grams 
of Rochelle salt dissolved in water and made up to 500 cc. 
These solutions should be preserved separately in rubber- 
stoppered bottles and mixed in equal volumes when needed 
for use. This is done to prevent deterioration. 

To 2 cc. of Benedict's solution in a test tube add 6 cc. 
of distilled water and not more than 7 to 9 drops of the 
urine under examination. Boil the mixture vigorously for 
15 to 30 seconds and allow it to cool to the room tempera- 
ture. If sugar is present in the solution a precipitate will 
form which is often bluish-green or green at first, especially 
if the percentage of sugar is low, and which usually becomes 
yellowish on standing. If the sugar present exceeds 0.06% 
this precipitate generally forms at or below the boiling 
point, whereas, if less than 0.06% of sugar is present the 
precipitate forms more slowly and generally only after the 
solution has cooled. 



39 



Benedict has further modified the test by making a 
single solution which does not deteriorate upon long standing. 
The formula is as follows : 

Cupric Sulphate 17.3 grams 

Sodium Citrate 173.0 " 

. Sodium Carbonate (anhydrous) 100.0 " 

Distilled water to 1000.0 cc. 

With the aid of heat the sodium citrate and carbonate 
are dissolved in about 600 cc. of water. Pour (through a 
folded filter if necessary) into a glass graduate and make 
up to 850 cc. The cupric sulphate is dissolved in about 
100 cc. of water and made up to 150 cc. The carbonate- 
citrate solution is poured into a large beaker and the cupric 
sulphate solution is added slowly, with constant stirring. 
The mixed solution is ready for use, and does not deteriorate 
upon long standing. 

The procedure is as follows: To 5 cc. of the reagent in 
a test tube add not more than 8 drops of the urine to be 
examined. The fluid is then boiled vigorously for one or 
two minutes and then allowed to cool spontaneously. In 
the presence of dextrose the entire body of the solution 
Will be filled with a precipitate, which may be red, yellow 
or green in color, depending upon the amount of the sugar 
present. If no dextrose is present, the solution will either 
remain perfectly clear, or will show a very faint turbidity, 
due to precipitated urates. 

Trommer's Test. To 4 or 5 cc. of urine, in a test tube, 
add one-half its volume of 20% caustic potash and 2 or 3 
drops of a solution of copper sulphate (1-10). Heat to 
the boiling point. If sugar be present a yellowish or red- 
dish precipitate is thrown down, the sugar having reduced 
the cupric hydrate to cuprous oxide. 

Bismuth Test. Put equal quantities of urine and 
20% solution of caustic potash in a test tube and add a 
pinch of subnitrate of bismuth. Boil the mixture and 
if glucose be present the powder turns black. Albumin 
and sulphur also reduce bismuth and must be removed if 
the test is to be reliable. Bismuth is more or less blackened 
with normal horse urine. 



40 



Nylander's Reagent. A solution is made of bismuth 
subnitrate 2 grams; Rochelle salts 4 grams; potassium 
hydroxide 10 grams; and distilled water to 100 cc. The 
urine is heated to boiling and a few drops of this alkaline 
solution of bismuth added, and on continuing the boiling, 
if sugar be present, the mixture turns black. The test is 
delicate, as little as 0.025% of glucose can be detected. 

Phenylhydrazine Test. (Modified from Kowarsky). 
To 5 drops of pure phenylhydrazine and 10 drops of glacial 
acetic acid in a test tube is added 1 cc. of a 10% solution 
of sodium chloride. After shaking the mixture, 3 cc. of 
the urine are added and the test tube heated for two minutes 
or longer. The fluid is then allowed to cool slowly. If 
the sugar content exceed 0.5% the precipitate of glucosa- 
zone takes place in about two minutes. Small quantities 
of albumin do not hinder the reaction, but are precipitated 
by boiling. After an hour or more examine the precipitate 
microscopically. A 10%, solution of sodium hydroxide 
has been recommended as a substitute for the sodium chloride 
solution and greater delicacy is claimed for the reaction. 

The glucosazone crystals will have the form typical 
of skeins of wool, which will generally be double, whereas 
other crystals precipitated, such as those of glycuronic 
acid, are irregularly formed. As little as 0.005% (one- 
fortieth grain per ounce) of glucose can be detected in urine 
by the phenylhydrazine reaction. 

Uric acid, urea, xanthine, and creatinin in no way 
stimulate the reaction of glucose with phenylhydrazine. 
The only bodies which can offer confusion are glycuronic 
acid and its derivatives. 

Fermentation Test. Robert's Differential Density 
Method. Take the specific gravity of the urine before adding 
the yeast and record it. Mix well 2 fluid ounces (60 cc.) of 
urine with yi cake of compressed yeast in a bottle. Set 
aside for 24 hours in a moderately warm place. After the 
fermentation filter and take the specific gravity again and 
subtract from that taken before Each degree of the remain- 
der represents one grain of glucose to the fluid ounce. Multi- 
ply by 0.219 to get the percentage. Thus: Specific gravity 
before fermentation, 1035; specific gravity after fermenta- 



41 



tion, 1015. 1035 — 1015 = 20 degrees of density lost, or 
20 grains of sugar to the fluid ounce. This test is con- 
clusive as to the presence of sugar, though it is not absolutely 
accurate as to quantity. 



VI. 

Bile in the Urine. In a number of pathologic conditions the 
elements of the bile are excreted in the urine. The bile pigments, 
bilirubin and biliverdin, may occur along with the bile salts, 
sodium glycocholate and taurocholate, or the bile salts alone may 
be present. 

Urine containing the bile pigments is colored a yellow brown 
or brownish green. It forms an intense yellow froth on agita- 
tion. It stains paper or linen a permanent yellow.* The bile 
pigments are found in jaundice, functional disorders of the liver 
(acute and chronic biliousness) ; organic diseases of the liver 
apart from jaundice (carcinoma, amyloid disease, cirrhosis) ; 
diseases of the spleen; fever; hemolytic diseases (anemia, leuco- 
cythemia and scurvy). 

Gmelin's Test. (Nitric acid containing nitrous acid).f 
Place a few drops of the suspected urine in a white porcelain 
dish and near them a few drops of the impure nitric acid; 
let the fluids run together and the usual play of colors is 
observed. 

Put some urine in a test tube and carefully pour in 
some of the yellow impure nitric acid, until it forms a stratum 
at the bottom. If the bile pigments are present at the 
line of junction of the fluids, a play of colors takes place — 
from above downwards — green, blue, violet or dirty red, 
and yellow. Nearly all urines give a play of colors, but 
green is the necessary and characteristic color to prove the 
presence of the bile pigments. 

Pettenkofer's Test for Bile Acids. Take about 2 cc. 
of suspected urine in a test tube and add 4 drops of a 10% 



*A little omnivorous or carnivorous bile may be added to the normal 
urine to demonstrate the tests if no icteric urine is available. 

fNitrous acid may be prepared by heating a little nitric acid to which 
a small amount of starch has been added. 



42 



solution of the cane sugar. Add strong sulphuric acid, 
drop by drop, cooling the tube in a dish of cold water im- 
mediately after adding the acid. Not more than 2 cc. of 
the acid should be used. Too much heat causes carboniza- 
tion of the sugar and the test is ruined. If bile acids are 
present, the fluid at first becomes opaque, then clear and 
successively brown, red and purple. It may require an 
hour or more to accomplish the test. This reaction depends 
upon the production of furfurol by the destruction of the 
sugar when the sulphuric acid is used. Furfurol in turn 
combines with cholalic acid, formed by the action of the 
sulphuric acid on the bile acids, giving the color. 

Pettenkofer's test may also be quite satisfactorily 
performed more quickly by putting a little of the suspected 
urine in a porcelain capsule, adding a few drops of a solution 
of cane sugar and then a few drops of strong sulphuric acid, 
keeping the mixture cool to prevent carbonization. 

Udranszky has modified Pettenkofer's test by using 
furfurol directly instead' of waiting for its formation by 
the action of the sulphuric acid upon the sugar. His pro- 
cedure is as follows: One cubic centimeter of the urine is 
treated with one drop of a 0.1% solution of furfurol and 
slowly superposed upon 1 cc. of sulphuric acid, care being 
taken to prevent too great heating of the mixture. A purple 
color appears at the plane of contact, that gradually extends 
upward into the superposed solution; on standing, the color 
turns bluish. In alcoholic solution a green fluorescence is 
seen. The pigment gives a typical spectrum. 

Hay's Test. Use two test tubes; in one place a little 
normal urine and in the other some of the suspected urine. 
Leave the test tubes in the rack in order to prevent agita- 
tion. Drop a little finely powdered sulphur upon the sur- 
face of the urines. If bile or biliary acids are present in 
the suspected urine the sulphur will sink at once to the 
bottom of the tube, while in the normal urine it will remain 
upon the surface or but a slight amount may sink, if the tubes 
are not agitated. 

Chloroform as a test for bile is quite satisfactory. Agi- 
tate a few drops of chloroform with the suspected urine in a 
test tube. If bile be present the chloroform becomes turbid 



43 



and acquires a yellowish hue, the depth of which is in propor- 
tion to the amount of bile present. 

To some of the suspected urine add a little bromine 
water (1%). If bile is present a green ring should appear. 
Blood. Blood is sometimes a constituent of the urine in 
disease. It may occur in two forms: 1. As hematuria, when the 
blood coloring matter is present in the urine in combination with 
blood corpuscles. 2. As hemoglobinuria, when no blood corpus- 
cles are present and the blood pigment or hemoglobin is in solution 
in the urine. 

Hematuria may have its source (1) in the kidneys due to 
injuries; acute nephritis; acute acerbation of chronic nephritis; 
diseases of renal vessels (embolism, thrombosis, aneurism, stasis); 
amyloid kidney (very rarely) ; infective fevers (smallpox, scar- 
latina, typhoid fever, etc.) ; certain blood diseases (scurvy, pur- 
pura, hemophilia); parasitic diseases (echinococcus) . 2. The 
renal pelvis and ureters due to renal calculi; tuberculosis; rup- 
ture of neighboring abscesses; parasites. 3. The bladder due to 
calculi;' cancer and other tumors; diphtheric cystitis; varicose 
veins; injuries. 4. The urethra due to injury (catheterization, 
impaction of calculi, etc.). 5. Extraneous discharges as the 
menstrual flow, etc. 

Hemoglobinuria has been observed in severe infectious dis- 
eases (typhoid fever, scarlatina, etc.) ; in conditions of blood 
dissolution (scurvy, purpura, etc.) ; in skin burns, sunstroke, etc. 
Heller's Blood Test is made by adding a little caustic 
soda solution to some urine in a test tube and heating. The 
precipitated phosphates are colored reddish brown and fall 
in a thick cloud to the bottom of the tube. 

Almen's Test. Add a few drops of freshly made 5% 
alcoholic tincture of guaiacum to the suspected urine. Shake 
well and add a few drops of hydrogen dioxide or of old oil of 
turpentine, and the mixture will turn greenish blue. The 
hemoglobin will change the color of the precipitate to blue. 
The test will not respond to a small quantity of blood. 

Hemin Test. If some blood or sediment supposed to 
contain blood be heated gently with some glacial acetic acid 
and a trace of sodium chloride, under a cover glass and 
then slowly evaporated in the air, brownish yellow rhombic 
crystals of hemin are found. The slide should be heated 



44 



two or three times, running a drop of the acid under the 
cover each time. 

The spectroscope and microscope are also used in blood 
tests. 

Melanine. In certain cases of melanosis this pigment 
appears in the urine, which, when emitted is clear, but gradually 
becomes of a deep brown, or even black color. 

Ordinarily melanine exists in solution in the urine, but some- 
times in the form of brownish or black sediment, recognizable 
by microscopic examination. Melanine may possess a diagnostic 
significance when the melanosis is beyond the reach of examina- 
tion by eye or touch. 

It may disappear from the urine when the disease is arrested, 
or it may remain stationary. Oxidizing agents, such as chromic 
acid and fuming nitric acid, transform the principle melanine, 
causing gradually with the first and immediately with the second 
a black coloration. According to Zeller, the most delicate test for 
melanine is bromine water. With melanine it gives at first a yel- 
low precipitate, which gradually blackens. Urobilin gives a yel- 
low precipitate with the same reagent, but it does not blacken. 

The practical significance of this condition (melanuria) is 
greatly limited by the fact that the urine may contain a large 
quantity of melanine in wasting diseases, whilst that derived 
from individuals suffering from melanotic cancer or sarcoma may 
be entirely free from it. Senator has recently confirmed this view 
by a series of clinical observations. Nevertheless, as an adjunct 
in diagnosis, the tests given are of undoubted utility. 



45 



VII. 



QUANTITATIVE ANALYSIS. 

Centrifugal Method. The centrifuge as ordinarily used gives 
the bulk percentage of the constituents, this percentage being 
arrived at arbitrarily as a result of numerous determinations 
upon normal urine. Knowing the normal bulk percentage it is 
not difficult to determine an abnormal or 
pathologic amount of a substance as indi- 
cated by its excess or deficiency, and the 
method is therefore a convenient and 
expeditious one for clinical purposes. 
To a beginner, however, the method as 
commonly employed, is misleading in 
that the bulk percentage and the true or 
actual percentages are widely different. 
Take for example the phosphates of the 
urine. The normal bulk percentage as 
given by the centrifuge is 8% for man; 
1% or less for the horse. Whereas, the 
real amount of phosphates present as 
P2O5 in the urine of man for the whole 
24 hours (1500 cc. of urine) is only 
about 3.5 grams, or in 1000 cc. about 2.5 
grams, or a true percentage of 0.25%. 
The true percentage may be calculated 
from the centrifuge by giving to each 0.1 cc. of precipitate a 
determined value with reference to the amount of V»Oi present. 

Each centrifuge tube is graduated to 15 cc. The first 10 cc. 
are divided so that each cubic centimeter is divided into 10 parts, 
each part representing 0.1 cc. The remaining 5 cc. are for hold- 
ing the test reagents which are added to the 10 cc. of urine. The 
first cubic centimeter because of the tapering end, will admit of 
finer graduation than the remainder of the tube. The first half 
of the cubic centimeter is divided into 1 /40 cc. (or .025 cc.) in 
order that small amounts of precipitate may be accurately read; 
the second half of the cubic centimeter is divided into 1 ,'20 cc. (or 
.05 cc.) ; while the remaining 9 cc. are each divided into 1 /10 cc. 
(or 0.1 cc.) as above stated. It is convenient to take the 1 /10 cc. 




Fig.9 

Hand Centrifuge. 



46 




Fig. 10. Aluminum Tubes. 



Percentage Sedimentation 
Tube. Tube. 



as the unit in the calculation of the true percentage. By taking 
the average of a great number of control tests with the burette it 
has been found that 1 ,/10 cc. of the precipitate, in the case of the 
phosphates, represents 0.13 gram of P 2 5 in 1000 cc. of urine. 
If albumin is present, remove it by adding a little acetic 
acid and applying heat. Filter and test the filtrate for the inor- 
ganic constituents. 

With the horse urine care must be exercised in adding the 
reagents. The large amount of carbonates present cause con- 
siderable effervescence, from the liberation of the CO2, and some 
of the urine is likely to flow over the tube. Add a little of the 
reagent and when the effervescence has ceased add a little more 
until the required amount has been used. 

Phosphates. Fill the graduated glass tube to the 10 cc- 
mark with the urine to be tested. Add 1 cc. of glacial 
acetic acid and 4 cc. of 5% uranium nitrate solution to 
reach the 15 cc. mark. Invert the tube several times and 
revolve in the centrifuge for three minutes. If, after revolv- 
ing three minutes at 1000 revolutions per minute, the preci- 
pitate comes up to the eighth 0.1 cc. line of the tube (0.8 cc.) 
multiply 0.13 by 8 for the product, which is 1.04 grams of 
P 2 5 in 1000 cc. If the total amount of urine for the 24 
hours is 1400 cc, the amount of P 2 O s present in this quantity 
can easily be calculated by the following proportion: 1.04 
gm. P 2 5 : 1000 cc. : : X : 1400. In which the value of X 
is found to be 1.456 grams of P 2 5 in 24 hours. Make the 
same determination with the urine of the horse. 



47 



Chlorides. Fill the graduated tube to the 10 cc. mark 
with the urine. Add 15 drops or 1 cc. of nitric acid to pre- 
vent precipitation of the phosphates. Then fill to the 15 cc. 
mark with the silver nitrate solution (1 to 8). Invert the 
tube several times to thoroughly mix the reagents. Revolve 
in the centrifuge for intervals of three minutes until the 
precipitate no longer settles. The average amount of preci- 
pitate for man is from 0.5 cc. to 0.8 cc; for the horse about 
0.5 cc. The value of each 0.1 cc. is 1.3 of a gram of NaCl 
per 1000. 

Sulphates. These are estimated as insoluble salts 
of barium. Fill the graduated tube to 10 cc. mark with 
fresh urine and add 5 cc. barium chloride solution, (4 parts 
barium chloride, 1 part hydrochloric acid, 16 parts distilled 
water) . 

The amount of precipitate for man and horse is about 
.075 cc. normally. The value of each 0.1 precipitate is 
2.4 grams of S0 3 . 

In the above tests write down in your notes the amount 
of each constituent for the 24 hours considering 1250 cc. 
as the total amount of urine passed. 

Quantitative Estimation of Uric Acid. This estimation is 
difficult and time-consuming and is generally regarded as inex- 
pedient for clinical purposes. The following methods are de- 
scribed because they are more suitable for clinical work although 
less accurate than the more elaborate methods of Salkowski, Lud- 
wig, Hopkins and others. 

Ruhemann's Uricometer for the rapid estimation of uric 
acid consists of a graduated tube with a glass stopper. It is a 
colorimetric method, in which iodine with carbon bisulphide serve 
as indicators. 

The following directions accompany each apparatus : 
Fill the glass tube to the lowest mark 5 with carbon bisul- 
phide. (It is not necessary for the tube to be quite dry, but 
there must be no drop of liquid at the bottom) . 

The lowest part of the convexity (double meniscus) has to 
be even with mark 5, see diagram. 

Add a solution consisting of iodine, 0.5 gm; potassium 
iodide, 1.25 gm.; absolute alcohol, 7.5 gms.; glycerine, 5 gms.; 
distilled water to make 100 gms. 



48 



» 



■ccm 
12.0- 
11.8 
1 1.6 - 
11.4 
11.2- 
ll.O-l 
10.8- 
10.6- 
10.4- 
.10.2- 
10.0- 
9.8- 
9.6- 
9,4- 
9.2- 
9.0- 
8,8- 
8.6- 
8.4- 
8,2- 
4,0- 
7.8- 
7.6- 
7.4- 
7.2- 
7.0- 
6.8. 
6,6. 
8.4- 
8.2 -HI 
6,0. 



6,6 

6.4. 

6.2. 

6,0- 

4.8. 

4.< 

4,4. 



8.6 
3.4 . 
3,2 
3,0 



0.175 
0.178 
0.181 
0.184 
0,187 
0.190 
0.198 
0.196 
0,199 
.0.202 
0,205 
0.208 
0,211 
0,215 
0.218 
0.221 
0.225 
0228 
.0.281 
0.235 
.0.238 
0,242 
0.245 
0.249 
0.252 
0.26 
.0.28 
.08 
0.33 
035 
0.88 
0.41 
.0,44 
0.47 
0,6 
0.65 
0.6 
0.653 
.0.7) 
-0.76 
-0.8 
.0,94 
.1.13 
--J.88 
Pl.63 
.1.89 
-215 
2.4(4 



J 

Fig. 11 

Ruhemann's 
Uricometer. 



Fill up so that the base of the upper arch 
of the double meniscus is on a level with mark 
J as shown in the illustration. 

Then add the urine to be examined, which 
has to be at a temperature of 18° Centigrade, 
to the mark of 2.45 (2.6 ccm.) 

Close the tube with glass stopper and 
shake well when the carbon bisulphide will be- 
come a dark copper brown color. 

After adding more urine under continued 
strong shaking, the carbon bisulphide will 
absorb all free iodine and the mixture will look 
like urine. 

Slowly adding more urine will change the 
yellow foam, created by the shaking, into 
white foam. 

The color of the carbon bisulphide will 
turn pink after a while. 

Should this color remain the same after 
the apparatus has' been shaken repeatedly and 
turned upside down, add another drop of 
urine and keep up the same procedure until 
only a slightly reddish coloration of the carbon 
bisulphide remains. 

Now shake again vigorously and the car- 
bon bisulphide will turn porcelain white and 
the urine will look like cloudy whey. 

To recapitulate: — The adding of urine 
has to be stopped as soon as the carbon bi- 
sulphide shows only a slightly reddish tint, 
because this will disappear entirely after re- 
peated shakings. The test is finished when the 
indicator appears snow-white, a sign that all 
iodine has been neutralized by the urine. 

To get rid of the remaining foam move 
the tube a few times slowly to a horizontal 
position, then open the stopper a little, to 
allow all liquid to settle in the tube. 

The proportion of uric acid is then read 
off the upper scale (per thousand of urine). 

The percentage is obtained by putting 



49 



a in front. If the upper meniscus line of the urine is between 
any of the 0.1 ccm. marks, the upper number should be read. 

The figures to the left of the apparatus refer to the number 
of cc. of urine added to the mixture. 

Should the urine contain less uric acid than the apparatus 
will in this way indicate, add the iodine solution to the mark half 
way between 5 and J and read after each reaction the half values. 

The vessel in which the urine is to be kept, must not be 
cleaned with soda. 

If the urine shows an acid reaction, it can be used at once, 
but if it should be alkaline, it has to be made acid by adding 
diluted acetic acid. Cloudiness is of no importance. 

If the urine contains a considerable sediment of sodium urate 
it should be well shaken. 

Strong colorations of the urine do not affect the action of 
carbon bisulphide. 

Traces of sugar and albumin do not disturb the result. 

If there is a very large percentage of albumin or traces of 
blood or pus, these pathologic substances have to be coagulated 
by boiling and the urine is filtered. 

The apparatus is not satisfactory for determining the uric 
acid in the urine of the horse. 

(The uricometer may be purchased of Eimer & Amend, New 
York). 

Estimation of Uric Acid by Weight. 20 cc. of hydrochloric acid 
are added to 200 cc. of urine and the mixture set aside for 24 hours. 
Uric acid crystals form and collect on the sides of the vessel. This 
may be collected on a weighted filter and washed thoroughly with 
water. The filter and uric acid are dried and weighed until there 
is no further loss of weight. The weight of the filter paper is deducted 
and the result gives the amount of uric acid in 200 cc of urine, from 
which the amount in 24 hours can be calculated. 

Urea. DoremUs Ureometer Test. Fill the long arm and 
bend of the ureometer with the hypobromite solution. (Hypo- 
bromite of sodium is prepared by mixing 2 cc. of bromine with 
23 cc. of a solution of caustic soda, 40 grams to 100 cc. of distilled 
water. To this mixture add an equal volume of distilled water). 
With a thoroughly washed pipette draw up exactly 1 cc. of urine 
and pass the pipette through the bulb of the ureometer as far as it 
will go in the bend. Compress the bulb of the pipette gently and 
steadily. The urine will rise through the hypobromite, and the 



50 



urea instantly decompose, giving off nitrogen gas. Withdraw 
the pipette after the urine has been expelled, taking care not to 
press the bulb hard enough to drive the air out with the urine, 
and read the volume of gas, after allowing the froth to subside. 

Each division mark on the ureometer indicates 0.001 gram 
of urea in 1 cc. of urine. The quantity of urea voided in 24 hours 
is ascertained by multiplying the result of the test by the number 
of cc. of urine passed during that period. 



rz\ 



/O. 









. Fig. 12 

Doremus 

Ureometer. 



Fig. 13. Hind's modification of 
Doremus Ureometer. 



The C0 2 resulting from the decomposition of the urea is 
absorbed by the excess of soda in the hypobromite solution, and 
nitrogen is evolved. 37.1 cc. of moist nitrogen gas measured 
under the ordinary conditions of experiment may be taken to 
represent 0.1 gram of urea. 1 gram of urea corresponds to 13.72 
grams of muscular tissue. 



51 



Albumin. Esbach's Method. Fill the tube with the 
suspected urine to the letter U, then add the reagent to the 
letter R. (The reagent is made by mixing 10 grams of picric 
acid and 20 grams of citric acid and adding 
enough water to make 1 liter. It is said that 
acetic acid may be substituted for the citric 
with as good results). Invert the tube a num- 
ber of times so that the contents may be 
thoroughly mixed. Close the tube tightly 
with the rubber stopper and set aside for 24 
hours, after which the amount of dn'ed albu- 
min in one liter of urine can be read in grams 
on the tube. The percentage is obtained by 
dividing by ten. Thus, if the coagulum stands 
at 3, the urine contains three parts of albumin 
per thousand, or 0.3%. If the albumin is 
very abundant (above four) the urine should 
be diluted to obtain an accurate result. Less 
than 0.5 per thousand of albumin cannot be 
accurately estimated by this method. 

Caution. The coagulable substance found in 
the urine of Bright's disease is a mixture of "serum-albumin" 
and "serum-globulin" or "para-globulin." Saturation of 
the urine with crystallized magnesium sulphate precipi- 
tates the serum-globulin, leaving the serum-albumin in 
solution. 

Centrifuge Test. Fill the graduated tube with urine 
up to the 10 cc. mark; add 5 cc. of Esbach's reagent. Invert 
the tube several times in order to thoroughly mix the fluids. 
Place the tube in the centrifuge and revolve for three minutes. 
Read off the amount of the precipitate; each 0.1 cc. is the 
equivalent of 0.21 of a gram of dry albumin per 1000 cc. of 
urine. 

Ferrocyanide Test with Centrifuge. The percentage 
tube is filled to the 10 cc. mark with the urine to be tested. 
Add 1 cc. of acetic acid and 4 cc. of a 5% solution of potassium 
ferrocyanide. Proceed as in the preceding test. Each 0.1 
cc. of precipitate is the equivalent of 0.21 gram of dry albumin 
per 1000 cc. of urine. 

Sugar. Einhorn's Fermentation Saccharometer. 



Fig. 14 

Esbach's 

Albumin- 

ometer 



52 




Fig. 15 

Einhorn's 

Saccharo- 

meter. 



Determine the specific gravity. Add 10 cc. of the sus- 
pected urine to 90 cc. of distilled water Put in a flask, 
add about one gram of compressed yeast and agitate 
thoroughly. Pour 10 cc. of the mixture into the 
bulb of the saccharometer and, by inclining the 
apparatus, the fluid will displace the air in the cylinder 
and remain there by atmospheric pressure. Be 
sure that no air bubbles remain in the cylinder. It 
is always well to test a normal urine at the same 
time and in the same way as a control. The mix- 
ture of normal urine with yeast will, on the following 
day, have only a small bubble on the top of the cylin- 
der. If, in the suspected urine, there is also present 
at the top of the cylinder a small bubble, no sugar 
is present ; but if there is a much larger volume of gas 
(C0 2 ) it is certain that the urine contains sugar. The 
apparatus should remain in a moderately warm place. As a 
result of fermentation the sugar is broken up into alcohol 
and C0 2 . The changed level of the fluid in the cylinder 
shows that the reaction has taken place and indicated by 
the numbers the approximate quantity of sugar present. 
The scale on the tube is empirical and indicates directly 
the percentage in the urine. 

Shieb's Test for Sugar in the Urine. 

Solution No. 1. 

Ammonium Sulphate (purest) 

Copper Sulphate (purest) 

Distilled Water 
Solution No. 2. 

Caustic Potash C. P. 

Distilled Water 

Dissolve and when cool, add 

Glycerine - 

Ammonia water 0.960 sp.gr. 

Add No. 1 to No. 2 and dilute the whole to 500 cc. 
with distilled water. Stopper securely and shake till thor- 
oughly mixed. 

Heat one dram of this solution in a test tube to boil- 
ing. Add the urine drop by drop, at slow intervals, boil- 



1.2 


grams 


2.6 


grams 


50. 


cc. 


20 


grams. 


50 


cc. 


50 


cc. 


300 


cc. 



53 



ing after each addition until the blue color has been dis- 
charged and the fluid has a light amber color or is colorless. 

If the solution is decolorized by 3 minims of urine it 
contains 9 to 10 grains of sugar per oz. 

If the solution is decolorized by 4 minims of urine it 
contains 7 to 8 grains of sugar per oz. 

If the solution is decolorized by 5 minims of urine it 
contains 5 to 6 grains of sugar per oz. 

If the solution is decolorized by 6 minims of urine it 
contains 4 grains of sugar per oz. 

If the solution is decolorized by 7 minims of urine it 
contains 3 grains of sugar per oz. 

If the solution is decolorized by 9 minims of urine it 
contains 2 grains of sugar per oz. 

If the solution is decolorized by 10 to 17 minims of 
urine it contains 1 grain of sugar per oz. 

If the urine contains more than 10 grains of sugar to 
the ounce it must be diluted with an equal quantity of water, 
and the number of grains per ounce multiplied by two. 

A very similar preparation, accurately compounded, 
is on the market under the name of Whitney's Reagent. 
It is for sale by the Norwood Chemical Co., 105 W. 40th 
St., New York. 

The following table gives the amounts of sugar in analyti- 
cal testing with Whitney's Reagent: 



If reduced by 


It contains to the oz. 


Percent 


1 minim 


16 grains or 


more 


3.33 


2 minims 


8 grains 




1.67 


3 " 


5.33 grains 




1.11. 


4 " 


4 




0.83 


5 " 


3.20 " 




0.67 


6 - " 


2.67 " 




0.56 


7 


2.29 " 




0.48 


8 " 


2 




0.42 


9 " 


1.78 " 




0.37 


10 " 


1.60 " 




0.33 




(1 




0.21) 



54 



BENEDICT'S QUANTITATIVE METHODS 

Copper Sulphate crystals - - 18 grams 

Sodium Carbonate - - 200 " 

Sodium Citrate - 200 " 

Potassium thiocyanate - - 125 " 

Potassium ferrocyanide (5% Sol.) 5 cc. 

Distilled water to - - - 1000 cc. 

Preparation: Dissolve the sodium citrate, sodium 
carbonate and potassium thiocyanate in 800 cc. of distilled 
water with the aid of heat and filter. Dissolve the copper 
sulphate separately in 100 cc. of distilled water and mix 
the two solutions slowly with constant stirring; then add 
the potassium ferrocyanide solution. Cool and dilute with 
distilled water to exactly 1000 cc. 

Application. To 25 cc. of Benedict's solution in a 
small beaker add from 4 grams to 5 grams of anhydrous 
sodium carbonate and heat the mixture to boiling over a 
wire gauze until the carbonate has been brought into solu- 
tion. Add a little distilled water occasionally to make up 
for evaporation. 

Place the urine under examination in a burette and 
run it into the hot Benedict Solution rather rapidly until 
the formation of a heavy chalk-White precipitate is noted 
and the blue color of the solution lessens perceptibly in its 
intensity. From this point in the determination from 2 
to 10 drops of the urine should be run rather slowly into 
the boiling Benedict Solution at one time, boiling the solu- 
tion vigorously for about 15 seconds after each addition. 
Complete reduction of the copper is indicated here as in 
Fehling's original method, by the complete disappearance 
of all blue color and the white precipitate of cuprous thiocya- 
nate. The end-point here, however, is very sharply defined, 
contrary to the conditions in the older method. 

To prevent the annoying bumping which often inter- 
feres with the titration, a medium-sized piece of washed 
absorbent cotton may be introduced into the solution. 
This cotton may be stirred about through the solution as 
the titration proceeds and the bumping thus eliminated. 
Calculation. Twenty-five cubic centimeters of Bene- 



55 



diet's Solution is completely reduced by 0.05 gram of dextrose. 
If Y represents the number of cubic centimeters of urine 
necessary to reduce the 25 cc. of the solution we have the 
following proportion Y : 0.05 : : 100 : X (percentage of 
dextrose) . 

Ehrlich's Diazo-reaction. This test has been recom- 
mended for the diagnosis of typhoid fever in man. It is 
stated that the reaction will also occur in some other dis- 
eases, but in spite of this fact the test is of value as an aid 
in the diagnosis. Two solutions are prepared as follows : 

1. 



Sulphanilic acid 


2 gms. 


Hydrochloric acid 


50 cc. 


Distilled water 


1000 cc. 



2. A. 0.5% solution of sodium nitrate. 

In performing the test, 50 parts of No. 1 and 1 part 
of No. 2 are mixed, and equal parts of this mixture and 
of the urine in a test tube are rendered strongly alkaline 
with ammonia. If the reaction be positive, the solution 
assumes a carmine-red color, which on shaking must also 
appear in the foam. Upon standing 24 hours a greenish 
precipitate is formed. The test must not .be considered 
positive unless a distinct coloration extends to and includes 
the foam on shaking. 



VIII. 
VOLUMETRIC METHODS. 



Chlorides. Mohr's method. Principle: If silver nitrate 
added to a solution containing sodium chloride, neutral potas- 
sium chromate, and an alkaline phosphate, the chloride is first 
precipitated, then the chromate, and lastly, the phosphate. The 
formation of the red silver chromate indicates the complete 
precipitation of the chloride: 

Solutions required: 

1. Standard solution of silver nitrate: 

Fused with silver nitrate 29.075 grams. 
Distilled water to make 1000 cc. 



56 



2. Saturated solution neutral potassium chromate: 
Neutral potassium chromate 10 grams. 
Distilled water to make 100 cc. 
Process. The urine should not be high colored, and 
should be free from albumin or excess of uric acid or mucus. 
Dilute 10 cc. of the urine with 100 cc. of distilled water. 
Fill a burette with the silver solution to the zero mark. 
Drop it slowly into the urine, stir it well and occasionally 
carry a drop of the mixture so as to come in contact with a 
little of the chromate solution in an evaporating dish. Test 
in this way until the first trace of orange color appears in the 
chromate solution. Make sure that the precipitation of the 
chlorides is complete by adding another drop of the silver 
solution from the burette. Read off the amount of silver 
solution used and calculate the result. 
Example : 

Quantity of urine in 24 hours, 1250 cc. 
Silver solution used, - 7.5 cc. 

1 cc. silver solution equals .01 gram NaCl 

.01 X 7.5 

X 1250 = 9.375 grams. 



10 

Make two or three determinations and take the aver- 
age for your final result. Do the same in the phosphate 
and sulphate tests. 

If the urine of the horse is very dark colored, it may 
be filtered through animal charcoal to make it light colored. 
Some of the chlorides may be held by the charcoal and thus 
diminish the amount in the urine tested; or the urine may 
be diluted with an equal volume of distilled water and the 
result multiplied by two. 

Gravimetric Method. 

A more accurate method is the following: If the urine is high 
colored, and contains albumin, or excess of uric acid and mucus, they 
must be removed. To do this, measure 10 cc. of the urine into a 
platinum capsule, add 2 grams of pure potassium nitrate, evaporate 
to dryness, and ignite at a dull red heat to destroy organic matter. 
When cool, treat the residue with hot water and filter. Acidulate 
the filtrate with dilute nitric acid, neutralize with carbonate of lime 
and proceed as above. 

Estimation of Phosphoric Acid. (Estimated as P2O5). By 
uranium acetate or nitrate. This method is based upon the fact 



that when a solution of acetate or nitrate of uranium is added to 
a solution containing soluble phosphates, sodic acetate and free 
acetic acid, all the phosphoric acid will be precipitated as phos- 
phate of uranium. This precipitate is of light yellow color, 
insoluble in acetic, but soluble in hydrochloric acid. The point 
of completion of the phosphoric reaction may be ascertained by 
placing with a glass rod a drop of the yellow mixture in contact 
with a drop of potassium ferrocyanide solution upon a white 
plate or filter paper. As soon as there is the slightest excess of 
the uranium solution after the phosphates have been satisfied, a 
brown precipitate will result in the mixture due to formation of 
ferrocyanide of uranium. The cochineal solution, noted below, 
serves as a better indicator for the horse urine than the ferro- 
cyanide. The following solutions are used: 

1. A Solution of Cochineal prepared by boiling 40 grams 

of cochineal in 800 cc. of water. When cool add 

200 cc. of alcohol and filter. 

Or a solution of potassium ferrocyanide. 

Potassium ferrocyanide, - 5 grams. 

Distilled water - 100 cc. 

2. Solution of Sodium Acetate. 

Sodium acetate, 100 grams. 

Acetic acid, - 100 cc. 

Distilled water to make 1000 cc. 

The following procedure is of use in standardizing the uranium 
solution. 

3. Standard Solution of Disodic Hydric Phosphate, made by dissolving 
10.0845 grams of the crystallized salt in water and diluting to 1 liter. Each 
cc. of this solution contains .002 of a gram of P2O5. In 5 cc. there is 0.1 gram 
of P2O5. 

4. Uranium Acetate. Since this cannot be obtained sufficiently pure 
to be weighed out and used directly we make a solution of it of indefinite 
strength and standardize it with other solutions. 

It has been found best to make the solution of uranic acetate of such 
a strength that each cc. will precipitate .005 of a gram of P2O5. Consequently 
every 2 cc. of the uranium acetate should be made equal to every 5 cc. of 
the sodic phosphate, or upon adding 20 cc. of the uranium acetate to 50 cc. 
of the sodium phosphate and then touching the plate or paper which has 
been moistened with the cochineal solution with the drop of the mixture, we 
should just get the green color. 

Put 50 cc. of the sodium phosphate with 5 cc. of the sodium acetate 
solution into a beaker. To this add slowly from the burette, the uranium 
acetate, testing occasionally, for the color on the paper. Suppose that on 
the addition of 8 cc. from the burette, the color is obtained, then 8 cc. of the 
uranium acetate are as strong as 20 should be, and for every 8 cc. of the 
uranium solution that we have, 12 cc. of water should be added. If it should 



58 

require more than 20 cc. to produce the color, the uranium solution must be 
concentrated by evaporation or more of the solid salt added. The solution 
has now been graduated or standardized. 

Application. 50 cc. of the clear urine, with 5 cc. of 
the sodium acetate solution are poured into a beaker and 
heated; to this the uranium acetate is slowly added from 
the burette. The mixture is constantly stirred with a glass 
rod, which should be applied frequently to the cochineal 
solution. As soon as the green color is obtained the process 
is completed. Read from the burette the amount of uranium 
acetate solution used. 

Example. Urine in 24 hours equals 1180 cc. Solution 
uranium acetate used equals 24.3 cc. 

.005 X 24.3 cc. 

X 1180 = 2.86 grams Po0 5 . 

50 

A shorter and approximately accurate method based 
on the above is as follows: To 10 cc. of the urine in a test 
tube add 1 cc. of sodium acetate and 1 cc. of the cochineal 
solution and boil. Add 5% uranium nitrate solution, 1 
minim at a time, boiling after each addition, until the mixture 
turns green. Each minim of the uranium solution represents 
0.046 gram of P 2 5 in 1000 cc. of urine. Multiply .046 by 
the number of minims required to produce the green color, 
and the product will represent the amount in grams of P2O5 
per liter in the given sample of urine. 

Estimation of Total Sulphuric Acid. (Estimated at S0 3 ). 
Dissolve 30.5 grams of pure crystallized chloride of barium in 
some distilled water and dilute to 1 liter. Each cc. of this solu- 
tion will equal .01 gram of SO3. A dilute solution of sodium 
or magnesium sulphate will also be required. 

Application. 50 cc. of clear urine are poured into a 
beaker, acidified quite strongly with hydrochloric acid, 
and heated over the flame. Let the solution boil for about 
ten minutes, the lamp is then removed and the barium 
chloride is allowed to flow slowly from the burette into 
the beaker and it must continue to flow as long as the preci- 
pitate is seen to increase. The precipitate is allowed to 
subside, then more of the barium chloride is added, and 
this process repeated, until no further precipitate is pro- 



59 



duced. Much time and labor will be saved by filtering a 
few drops of the solution now and then, and allowing these 
to drop into the test tube containing some of the dilute 
sodium or magnesium sulphate. As soon as an excess of 
the barium chloride has been added, a precipitate will appear 
in the test tube. Read from the burette the amount of 
barium chloride used; each cc. of which will indicate .01 
of a gram of SO3 in each cc. of urine, and from this the total 
amount may be calculated. 

Example: Urine 2000 cc. Amount of barium chloride 
solution 25 cc. 

.01 X 25 

X 2000 = 10 grams S0 3 . 

50 

Relation of Urinary Constituents in Normal Human Urine. 

There is normally a direct proportion within narrow limits, between 
the solid substances of the urine, which it is desirable to keep in 
mind. 

Relation of Urea to total solids 50% or one-half. 

" inorganic matter to other solids 30%. 

" uric acid to urea 2.5% or 1 /40. 
' nitrogen in urea to total nitrogen 91%. 

" phosphoric acid to urea 12.5% or }/g. 

" chloride of sodium to urea 40%. 

" the sulphates to total nitrogen 18%. 



IX. 
CHEMICAL EXAMINATION OF URINARY DEPOSITS. 

There is generally a more or less voluminous deposit in the 
urine after standing for 24 hours; sometimes this deposit is 
formed in the bladder and sometimes it forms after the emis- 
sion of the urine; it is important to note this fact. Deposits or 
sediments should not be confounded with calculi. The former 
have a pulverunt form while the latter have the appearance of 
grains or granules of greater or less size. 

It is often very easy to determine the nature of the deposit 



60 



by a microscopic examination, but chemical reagents in some 
cases give more precise results. 

With the urine containing sediment, shake thoroughly to 
distribute it, then pour into a centrifugal tube and revolve until 
the sediment is completely precipitated. Note the reaction of 
the urine, whether acid or alkaline. Pour off the clear superna- 
tant fluid. If the urine is acid pursue the following scheme. 

Acid Urate of Soda. If the deposit is more or less red, treat 
with a little boiling water. If the sediment is dissolved it is com- 





o 

dfoQ 







Plate II. 

Various forms of Calcium Carbonate Crystals. (Horse Urine) 



61 



posed of the acid urate of soda. Caustic potash does not dissolve 
it, hydrochloric acid gives a crystalline precipitate, the deposit 
will give the murexide test. (Page 26). 

Uric Acid. If a crystalline deposit, almost insoluble in 
boiling water, but soluble in caustic potash and giving the murexide 
test, it is uric acid. 

Cystine (very rare). The deposit of crystalline, insoluble 
in caustic potash but soluble in ammonia and hydrochloric acid. 

Pus. A white, dense, mucus deposit, viscid and not often 
mixing with the urine; caustic potash renders it more viscid; 
the microscope shows the pus corpuscles. This deposit is rare 
in acid urines. 

Blood. Red deposit; the tincture of guaiac with hydrogen 
dioxide gives a blue color. The spectroscope gives the charac- 
teristic lines in the spectrum. The hemin test will show the 
characteristic crystals. When blood is present the urine is gener- 
ally albuminous. 

Some deposits not very abundant and having no special 
chemical reaction may be recognized under the microscope. 

Neutral or Alkaline Urine. 

Alkaline Urates. More or less reddish deposits easily soluble 
in boiling water and giving the murexide test. 

Triple Phosphate or Calcium Phosphate. A white deposit, 



B 







I 


;,,,,, 
















■ i l ' : M 










111 






I 




! hi° 




ORANGE YELLOW GiREEN BLUE VIOLET 




1 1 

ii 




II 


I 




I 



Fig. 16 

A. — Spectrum of Oxy-hemoglobin. B. — Spectrum of Reduced Hemo- 
globin. C. — Spectrum of Urobilin. 



62 



insoluble in boiling water, soluble in acetic acid, does not give 
the murexide test; acidified with nitric acid, the molybdate of 
ammonia solution gives a yellowish precipitate in the cold. 

Calcium Oxalate. White deposit insoluble in boiling water, 
also in caustic potash and acetic acid. Soluble in hydrochloric 
acid or in nitric acid; does not give the murexide test. 

C ale -urn Carbona'e. White deposit insoluble in boiling 
water or in caustic potash. Soluble with effervescence in acetic, 
nitric or hydrochloric acids, does not give murexide reaction. 

Pus, (more frequent in alkaline and albuminous urine). A 
white, dense, mucus-like deposit, not mixing readily with the rest 
of the urine. Caustic potash renders it more viscid, the micro- 
scope shows pus globules. 

Blood. Determined by the guaiacum test, spectroscope, 
microscope, or hemin test. 

From the pathologic point of view the presence of urinary 
sediments generally indicate an alteration of the secretions. 



X. 

Microscopical Examination of Urine. If an immediate 

examination is desired the centrifuge may be used to cause the 

sediment to separate from 

the urine; otherwise the 

urine is set aside for some 

hours in a cool place when 

the sediment gradually 

settles to the bottom. The 

supernatant liquid is poured 

off and by means of a 

pipette, camel's hair brush 

or a wire loop a small 

portion of the deposit is 
Fig. 17. Uric Acid Crystals. , . . ... , 

transferred to a slide and 

examined. 

The deposits may be divided into unorganized and organized 

sediments. 




63 



Unorganized Sediments. In unorganized sediments two con- 
ditions may be considered: 1st, the urine is acid. 2nd, the urine 
is alkaline. 

In acid urine look first for crystals of uric acid diverse in 
form and reddish brown in color. (Fig. 17). Second, crystals of 
acid urate of soda, yellowish or red. The crystals are badly 
formed. Third, crystals of oxalate of lime also found in alkaline 
urine. (Fig. 18). Fourth, crystals of hippuric acid. Fifth, 
crystals of calcium sulphate. (4 and 5 are met with rarely in 
acid urine). Sixth, calcium phos- 
phate. Make sure of the identifi- 
cation by reference to charts. 

In alkaline urine look first for 
ammonio-magnesium phosphates, 
(Fig. 19), (triple phosphates). 
Second, bicalcium phosphate crys- 
tals. Third, tricalcium or amorph- 
ous phosphate crystals. Fourth, 
crystals of sulphate of lime. Fifth, 
crystals of oxalate of lime, (also 
found in acid urine). Sixth, 

crystals of urate of ammonium in the form of yellow colored 
spheres. 

Crystals of cystin, leucin and tyrosin are sometimes en- 
countered in acid or alkaline urines. 

^Organized Sediments. These are brought more plainly into 




Fig. 18. Calcium Oxalate. 




Fig. 19. Triple Phosphates. 



04 




Fig. 20 

Micrococcus Ureae. 



view if the preparations are stained, although this is not uni- 
versally practised. Organized sediments may be divided into (a) 
histologic elements, (b) microbic elements. 

Of the histologic elements there are frequently encountered, 
1st, epithelial cells from the bladder, from the vagina and from 
the ureter, their presence has no special significance. If, how- 
ever, they are present in considerable quantity, a lesion of these 
parts may be suspected. 2nd, cells from the pelvis of the kidney, 
generally an indication of renal affection. 3rd, cancer cells, 
which have a special significance. 4th, 
hyaline or granular casts, seen more 
easily in stained preparations; they are 
encountered frequently in albuminous 
urine; their presence indicates a mild 
form of nephritis. 5th, epithelial hem- 
orrhagic or wax casts, of which the 
hemorrhagic are the most frequent, they 
are markedly colored and easily recog- 
nized on account of the hemoglobin they 
contain; they contain fine granulations 
which are not blood corpuscles but possible fragments of them, 
they indicate a severe form of nephritis. 6th, cylindroid ele- 
ments drawn out in an irregular and somewhat ribbon-like form; 
they are the product of the secretion of the urinary epithelium. 
In the human race they may be found in cases of scarlatina and 
generally in certain forms of nephritis. In examining for casts 
it is desirable to examine the urine immediately after emission, 
as they generally disintegrate rapidly and disappear. 7th, 
blood corpuscles 
more or less cre- 
nated but easily 
recognizable b y 
their yellowish 
tint; the presence 
of blood corpuscles 
in the urine indi- 
cates a hemorrhage 
either in the blad- 
der or in the kidney, 
borders, granular contents and appear quite refractive, a drop of 




Fig. 21. Red blood corpuscles. A. Normal. 
8th, pus corpuscles which may have crenated 



Go 



dilute acetic acid will render the nuclei visible. Pathologically the 
pus corpuscles indicate a suppuration of some portion of the 
urinary tract. In the majority of cases it is impossible to say 
if the pus comes from the. bladder or kidney. In cases of cystitis 
of the neck of the bladder, the last portion of the urine passed 
contains no pus, while if this trouble be in the kidney this last 
portion will contain pus. This fact is of some use in diagnosing 
cystitis of the neck of the bladder. 9th, spermatozoa may be 
present but are easily 
recognized by their 
elongated form. 10th, 
mucus may be fre- 
quently present, but 

has no very great C^SIfo" jEbJ2fa%& i&^ti 
p a t h o 1 o g i c signi- 
ficance. 




Fig. 22. Red blood corpuscles crenated. 



Glycogen Cells. If 

the urinary sediment 
is stained with dilute 
L u g o 1 ' s Solution, 
there will be seen, in 
many specimens of 
human urine, a num- 
ber of epithelial cells 
present with more or 

less of their interior stained a deeper brown like glycogen, than 
in other cells that may be present. As yet, nothing has been 
determined in regard to their importance. 

Amyloid or Amylaceous Bodies. These are small circular 
or oval bodies which give the starch reaction with an iodine solu- 
tion. Just what their significance is, is not known. Virchow first 
described them. They have been found in various tissues and 
excretions, including the urine normal as well as pathological. 
In diseases of the kidneys, Veitz and Wederhake found that these 
amyloids afford us important indications; but that in affections 
of the bladder they are present in increased quantity. Weder- 
hake therefore suggests that the amyloid bodies have a "certain 
value in differential diagnosis. The absence of the bodies in 
pathological urine is against catarrhal disease of the bladder; 
whilst their presence, especially if numerous, in urine which 



of)*** 




B 











G 



Jl 







i 









fe. 



1 






<^<* 



^ 



ft ^ 












«#' ft 











*> si/,. * '■ J&f-. \s£\ 




Plate III 

Urinary Sediment. A. Uric Acid. B. Ammonium Urate. C. Acid 
Sodium Urate. D. Urea Nitrate. E. (1) Leucin and (2) Tyrosin. F. 
Cystin. G. Ammonio- Magnesium or triple phosphate. H. Calcium Phos- 
phate. I. Calcium Oxalate. J. Blood Corpuscles. K. Pus and Mucus. L. 
Hemin Crystals. M. (1) Hyaline Casts, (2) Granular Casts. N. Epi- 
thelial Casts and Cells. O. (1) Waxy Casts; (2) Casts with Blood Cor- 
puscles; (3) Casts with Fat Globules. (After Simon). 



67 



appears to contain renal elements only, indicates that the bladder 
is also affected. (Dixon Mann). 

The urinary sediment stained with a dilute Lugol's solution 
show these bodies, when present, varying in color from a blue to 
a deep blue black color. 

Urinary Casts. Although probably observed earlier, Henle 
is credited in 1842, with first carefully describing the casts moulded 
in the tubules of the kidneys. Among earlier views, casts were 
considered as being composed of coagulated fibrin; as pro- 
ducts of the secretion of the epithelium of the tubules; as trans- 
formed or disintegrated epithelial cells and as products from 
the blood. A view quite commonly accepted is that casts are 
the products of the coagulation of albuminous material. The 
fact that the presence of casts in the urine is usually accompanied 
by the presence of albumin lends 
force to this view; for the more 
abundant the albumin, the more 
likelihood is there of finding casts. 
From this standpoint, then, casts, 
may be regarded as albuminous 
exudates from the blood, with the 
addition of transformed or de- 
stroyed epithelium. In nearly all 
cases where casts are present, some 
albumin is found. Occasion- 

ally they are sometimes described 
as being present without the ap- 
pearance of albumin. This condi- 
tion is looked upon with some 
doubt; for it is commonly be- 
lieved that albumin exists but in 
an amount too slight to be detected by the ordinary chemical 
tests. 

The presence of casts in the urine is of much diagnostic im- 
portance; if found in any quantity, they indicate nephritis par- 
ticularly if albumin is also present in any amount. It is claimed 
by some that merely hyperemia of the kidney will cause the 
appearance of casts in the urine, and that they may sometimes be 
found when the kidneys are perfectly intact. Mitchell states 
that two or three hyaline or one or two small granular casts may 




Fig. 23. a. Showing forma- 
tion of hyaline cast in tubule. 
b. Hyaline cast with granular 
deposit, c. Granular cast. 



68 



be found in one of every three specimens of the twenty-four 
hour's urine examined (human). Casts have been described in 
cases of gastro-intestinal catarrh, in jaundice, in acute and chronic 
anemia, as well as in nervous affections of different kinds, with- 
out accompanying inflammation of the kidneys. There are 
many, however, who hold that casts are always the products of 
an inflammatory process, and are, therefore, indicative of renal 
inflammation. Other evidences or symptoms should also be 

taken into account. 

Occasionally 
it is difficult 
to find casts 
even when 
they are 
known to 
exist. Alka- 
line urine has 
a tendency to 
dissolve them. 
At t im e s 
they will not 
settle for 
hours. Usu- 
ally if al- 
lowed to 
stand six 
hours, the 
casts will set- 
tle if they are 
present. The 
centrifuge will 

bring them down in' a few minutes. A low power of the micro- 
scope (150-200 diameters) may be used in the search for casts 
and will enable the observer to pass over the field quite rapidly. 
To identify the cast and its structure, a power of 400-500 diame- 
ters is required. More than one specimen must always be examin- 
ed before giving a positive statement as to the presence or 
absence of casts. False or pseudo-casts have been described; 
these are believed to be accidental formations, while true casts in 
general indicate nephritis. 




Fig. 24. a. Blood cast and hyaline cast carrying 
blood cells, b. Pus cast. c. Hyaline cast carrying epi- 
thelial cells, d. Epithelial casts. (Greene). 



69 



True casts may appear in three different sizes according to 
the portion of the tubule in which they are formed. The smaller 
size originates in the narrow tubule. The next in size comes from 
the convoluted tubule of the second order (no casts from the 
convoluted tubule of the first order, i. e., that portion of the 
tubule nearest the glomerules, appear in the urine, because they 
are too large to pass through the narrow tubules). The third and 
largest are those developing in the straight collecting tubules. 
It is believed a prognostic value may be attached to the size of 
the casts as well as to their number. Casts from the narrow 
tubules indicate a mild attack; from the convoluted tubules a 
severe form, especially in the cortex of the kidney; from the 
collecting tubules, with the other forms also present, a serious 
condition of general renal inflammation or unfavorable prognosis. 
The identity of casts may generally be determined by their uni- 
form width. They are usually longer than they are broad, and 
have one well- 
rounded ex- 
tremity and 
well defined 
borders. 

True casts 
are of six va- 
rieties ; hya- 
line, epitheli- 
al, blood, 
granular, fat- 
ty and waxy 
casts. In a 
general way 
the first three 
varities are 
found in an 
acute form of 
nephritis . 
During the 

first few weeks of the inflammation the last three varieties are not 
encountered. If the acute condition passes into a subacute, then 
the granular variety appears, at first in small number, then in 
larger, with, usually, a considerable number of the hyaline and 




Fig. 25. Granular Casts. (Greene) 



70 



epithelial casts. Fatty and waxy casts are always secondary 
products, and as a rule not found until a nephritis has existed 
for some time. 

Hyaline casts are colorless, pale, more or less transparent 
formations soluble in acetic acid. They are of variable size and 
generally difficult to detect on account of their apparently struc- 
tureless character. At times a slight granulation may be seen 
imbedded in or adhering to their matrix and occasionally acci- 
dental attachments of pus or fat globules in small numbers. 

Epithelial casts 
have a hyaline ma- 
trix more or less 
concealed by epi- 
thelial cells. The 
presence of these 
casts is indicative 
of an acute process. 
Blood casts con- 
sist of the hyaline 
matrix with blood 
corpuscles imbed- 
ded in or adhering 
to the matrix. Pus 
casts are rare, but 
when present the 
pus corpuscles ad- 
here to the matrix. 
Blood casts are indicative of a hemorrhage into ■ the tubules and 
of an acute hemorrhagic process. Hyaline and epithelial casts 
are usually associated with them. 

Granular casts usually 
have well denned boundaries 
with granular matter imbed- 
ded in or adhering to the 
matrix. They may be finely 
or coarsely granular, the lat- 
ter having a more serious 
significance. Granular casts 
are due to a disintegration of 
the renal epithelium. Their Fig. 27. Fatty Casts and Fat Droplets. 




Fig. 26. a. Fatty casts, b. and c. Blood 
casts, d. Free fatty Molecules. (Roberts). 




1 




degree of refraction is changeable; sometimes they appear yel- 
lowish, at other times colorless. 

Fatty casts have a hyaline matrix containing a number of 
small, glistening fat globules and granules. Some free fat is 
also usually found in the field. As fatty casts are secondary 
products of epithelial and granular casts, the diagnosis of a 
chronic process is justifiable. The hyaline matrix is character- 
istic of the different varie- 
ties of casts that have 
been mentioned up to this 
point. 

Waxy casts differ in 
chemical composition from 
those previously m e n - 
tioned. They are charac- 
terized by wavy contours; 
a high refracting power; a 
more or less yellowish color 
and quite a high degree 
of brittleness. They are 
slowly, if at all, attacked 
by acetic acid. Their presence signifies waxy degeneration 
of the kidney. Hyaline casts may sometimes have a superficial 
resemblance to waxy casts, but they never have the same high 
refraction as the latter. 

In the urine of the horse, the sediment of crystals of lime car- 
bonate may obscure the search for casts. In this case add a few 
drops of acetic acid which will dissolve the crystals quickly, 
and the casts, if present, will show more distinctly but undergo 
solution more slowly than the crystals. 

Cast-like formations are composed of various elements having 
a form somewhat similar to casts but lacking the matrix soluble 
in acetic acid. Amorphous urates often simulate granular casts 
in form. Bacteria are often grouped in a manner similar to the 
form of cast, but a close inspection shows an irregular outline, 
and usually a number of groupings not in cast form. Granular 
detritus and hematoidin may also assume the form of casts; like- 
wise epithelial cells, blood corpuscles and fibrin in renal hemor- 
rhages may also assume the form of casts. Acetic acid is said to 
be a reliable reagent for differentiating between true and false 



Fig. 28. Waxy Casts. 



72 



casts. This reagent dissolves the matrix of the true casts but does 

not act upon the cast-like formations. 

Cylindroids appear like hyaline casts, but are large and 

band-like. Their breadth is uniform and they often contain 

crystals, epithelial cells and corpuscles. 
They are soluble in acetic acid and 
are of renal origin. No special sig- 
nificance is attached to them. 

Mucous cylinders sometimes call- 
ed cylindroids are usually not of 
uniform breadth and seldom contain 
morphologic constituents and are in- 
soluble in acetic acid. They are usu- 
ally found in any urine containing 
an abundance of mucus and are of 
no special significance. 




Fig. 29. Cylindroids. 

a. b, Cast-like forms; c. 

Filamentous forms. (Ogden). 



Upon special blanks, test specimens of urine quantitatively, 
the number and amount of constituents being unknown. 



Form used in examination of Horse Urine. 

Owner Address 



No Species 

Object 



Age . . . 
. Date 



Sex 



Amount in 24 hours 

Specific Gravity 

Reaction 

Color 

Translucency 

Consistency 

Total Solids 

Chlorides 

Sulphates 

Phosphates 

Urea 

Uric Acid 

Hippuric Acid 

Indican 



Normal (Horse). 
3000-4000 c. c. 
1025-1050 
Alkaline 

Yellowish brown 
Turbid 
Viscid 
50-120 parts per 1000 

8-14 " 

2-3 

.05-.2 " 
20-40 " 
Trace 

4- 8 parts per 1000 

.1-.2 " 



Sample 



ABNORMAL CONSTITUENTS. 

Albumin 
Sugar 
Bile 
Hemoglobin 

MICROSCOPIC EXAMINATION. 
Epithelial Cells 
Leucocytes 
Blood 
Casts 

Spermatozoa 
Micro-organisms 

Crystalline Deposit. 
Calcium Carbonate 
Calcium Oxalate 
Triple Phosphates 



74 



Form used in Examination of Human Urine. 



Name 




Address 


Date. 


Normal. Sample. 


Amount in 24 hours. 


1000-1500 cc. 


Specific gravity 


1015-1025 


Reaction 


Acid (30°-40°) 


Color 


Light golden 


Translucency 


Clear 


Sol'ds 


34-50 parts per 1000 




50-75 parts per 24 hours 


Chlorides 


6-9 parts per 1000 as NaCl 




10-15 parts per 24 hours 




1.5-2.5 parts per 1000 as P 2 5 


Phosphates 


2.-3.5 parts per 24 hours 




1.5-3 parts per 1000 as SO 3 


Sulphates 


2.5-4 parts per 24 hours 


Urea 


14-22 parts per 1000 




22-33 parts per 24 hours 


Uric Acid 


0.25-0.40 parts per 1000 




0.40-0.60 parts per 24 hours 


Indican 


Normal, little, medium, much 


Albumin 




Sugar 




Bile 




Hemoglobin 




MICROSCOPIC EXAMINATION. 


Epithelial cells 




Leucocytes 




Blood 




Casts 


• 




Crystalline Deposit. 


Uric Acid 




Oxalates 




Triple Phosphates 


• 


Urates 





70 



Procedure in Examining a Sample of Urine. 

Experience has shown that the following procedure expedites 
the work of examination. Pour the urine into a sedimenta- 
tion glass, as some little time may be required for the sediment 
to settle. After noting the amount, color and translucency, 
take the specific gravity with the urinometer. This may be 
done in the sedimentation glass. Filter as much of the urine 
as may be needed for the subsequent tests. While waiting for 
the urine to filter make the tests for urea and uric acid as the 
correct reading is not obtained until some few minutes after 
the tests have been made. Test the reaction with litmus paper 
or if the degree of acidity is required (human) use the acidi- 
meter. Test for indican. Thus far the tests may be made just as 
well with the unfiltered as the filtered urine. 

For the albumin and sugar tests the filtered urine must 
be used, using two or three different tests for each substance. 
If albumen is present it should be removed before making the 
sugar tests. Also before making the centrifuge tests for the 
chlorides, phosphates and sulphates. This may be done by adding 
a little acetic acid to the urine and boiling it until the albumin 
is coagulated. Filter out the albumin and use the urine thus 
filtered for the remaining tests. 

With a pipette remove some of the sediment at the bottom 
of the sedimentation glass and introduce it into a centrifuge 
tube filling the remainder of the tube to the required height 
with the unfiltered urine. Include this with the other centri- 
fuge tubes used in making the tests for the chlorides, etc., revolv- 
ing them in the centrifuge for three minutes. If albumin is 
present and it is desired to know the amount, it may be deter- 
mined by the centrifugal method as described in the text. 

For the microscopial examination, prepare a couple of slides 
from the sediment in the centrifuge tube, after pouring off the 
clear urine above. It is well, also, to prepare a slide 'or two 
from the sediment in the sedimentation glass. After putting 
a drop or two of the sediment on the slide it may be covered with- 
a cover glass and examined clear or before covering the sedi- 
ment a drop of dilute Lugol's solution, or safranin or other stain 
may be added. In some cases the slight tint of the dye appears 
to make the casts or other elements stand out more clearly. 
To detect the amyloid bodies or glycogen cells it is necessary 



76 



to use the iodine solution and this often times serves to make 
the casts and other elements more distinctly visible. 



APPENDIX. 
Formulae for Reagents. 

Barium Chloride Solution for Centrifuge Sulphate Test. 
Barium chloride, 4 parts 

Hydrochloric acid, 1 part 

Distilled water, 16 parts 

Barium Chloride Solution for Quantitative Test of Sulphates. 

Pure crystallized barium chloride, 30.5 grams. 

Distilled water to 1000. cc. 

Baryta Mixture. This mixture is prepared by making sat- 
urated solutions in the cold, of barium nitrate and barium hydrate, 
and adding two volumes of the hydrate to one volume of the 

nitrate. 



Benedict's Solution. 






Solution 1 . Cupric Sulphate 


34.65 


grams 


Distilled water up to 


500. 


cc. 


Solution 2. Sodium carbonate (anhy-' 






drous) , 


100. 


grams 


Rochelle salt, 


173. 


grams 


Distilled water up to 


500. 


cc. 


Benedict's Single Solution. 






Cupric Sulphate, 


17.3 


grams 


Sodium Citrate, 


173. 


grams 


Sodium Carbonate (anhydrous), 


100. 


grams 


Distilled water up to 


1000. 


cc. 


Benedict's Solution for Quantitative Tests. 




Copper Sulphate Crystals, 


18 grams 


Sodium Carbonate Crystals, 


200 grams 


Sodium Citrate, 


200 grams 


Potassium Thiocyanate, 


125 grams 


Potassium Ferrocyanide (5 %Sol.) 


5 cc 




Distilled water to 


1000 cc 





Dissolve the citrate, carbonate and thiocyanate in 800 cc. 



77 



of distilled water with the aid of heat. Dissolve the copper 
sulphate separately in 100 cc. of distilled water and mix the two 
solutions slowly with constant stirring ; then add the f errocyanide 
solution. Cool and dilute with distilled water to exactly 1000 cc. 

Cochineal Solution. Boil 40 grams of cochineal in 800 cc. 
of water. When cool add 200 cc. of alcohol and filter. 

Decinormal Oxalic Solution. Dissolve 6.285 grams of pure 
oxalic acid in enough distilled water to make at or near 15° C. 
(59° P.), exactly 1000 cc. 

Decinormal Sodium Hydroxide Solution. Dissolve 3.996 
grams of pure sodium hydroxide in enough distilled water to 
make, at or near 15° C. (59° F.), exactly 1000 cc. 

Ehrlich's Diazo-reaction. 

Solution 1. Sulphanilic acid, 2 grams 

Hydrochloric acid, 50 cc. 

Distilled water, 1000 cc. 

Solution 2. A 0.5% solution of sodium nitrite. Use in the 
proportion of 1 part of No. 2 to 50 parts of No. 1. 

Esbach's Reagent. 

Picric acid, 10 grams 

Citric acid, 20 grams 

Distilled water up to 1000 cc 

Fehling's Solution. 

Solution A. Pure Copper Sulphate crystalline. 34.64 grams 
Distilled water up to 500. cc. 

Solution B. Sodio-potassium tartrate (Rochelle 

salt), 173. grams 

Pure caustic potash, 125. grams 

Distilled water up to 500. cc. 

Use equal parts of A and B . 

Hypobromite Solution. Dissolve 40 grams of caustic soda 
in 100 cc. of distilled water. To 23 cc. of this solution add 2 cc. 
of bromine. To this mixture add an equal volume (25 cc.) of 
water. The solution is not very stable and is best made up as 
needed. 



78 



Lugol's Solution. 

Iodine, 2.5 grams 

Potassium iodide, 5. grams 

Distilled water to 50. cc. 

This may ordinarily be diluted 1 to 5 or 10 for urine work. 

Magnesia Mixture. 

Magnesium sulphate, 1 part. 

Ammonium chloride, 1 part 

Ammonia water, 1 part 

Distilled water, 8 parts 

Millard-Roberts Reagent. 

Nitric acid, 1 part 

Sat. Sol. Magnesium Sulphate, 5 parts 

Mohr's Volumetric Method for Chlorides. 

Solution 1. Fused silver nitrate, 29.075 grams 

Distilled water to make, 1000. cc. 

Solution 2. Neutral potassium chromate, 10. grams 
Distilled water to make, 100. cc. 

Nylander's Reagent. Digest 2 grams of bismuth subnitrate 
and 4 grams of Rochelle salt in 100 cc. of a 10% solution of potas- 
sium hydroxide. The reagent should then be cooled and filtered. 

Obermayer's Reagent. Dissolve 2 grams of solid ferric 
chloride in 500 cc. of concentrated hydrochloric acid. Sp. gr. 1.19. 

Phenolphthalein. Dissolve 1 gram of phenolphthalein in 
100 cc. of 95% alcohol. 

Ruhemann's Uric Acid Reagent. 

Iodine, 0. 5 grams 

Potassium iodide, 1.25 grams 

Absolute alcohol, 7.5 grams 

Glycerine, 5. grams 

Distilled water to 100. grams 

Salicylic Adehyde. 

Dissolve 1 gram of salicylous acid in 10 cc. of absolute alcohol. 
Shieb's Reagent. 

Solution 1. Ammonium Sulphate (purest), 1.2 grams 
Copper Sulphate (purest), 2.0 grams 

Distilled water up to 50. cc. 



79 



Solution 2. Caustic potash C. P. 
Distilled water up to 
Dissolve and when cool, add 
Glycerine, 
Ammonia water, 0.960 sp. gr. 

Add No. 1 to No. 2 and dilute the whole. to 500 cc. with 
distilled water. Stopper securely and shake until thoroughly 
mixed. 



20. 


grams 


50. 


cc. 


50. 


cc. 


300. 


cc. 



